U.S. patent application number 13/909013 was filed with the patent office on 2014-01-30 for influenza virus reassortment.
This patent application is currently assigned to Novartis AG. Invention is credited to Philip DORMITZER, Bjoern KEINER, Peter MASON, Pirada SUPHAPHIPHAT, Heidi TRUSHEIM.
Application Number | 20140030291 13/909013 |
Document ID | / |
Family ID | 47997362 |
Filed Date | 2014-01-30 |
United States Patent
Application |
20140030291 |
Kind Code |
A1 |
SUPHAPHIPHAT; Pirada ; et
al. |
January 30, 2014 |
INFLUENZA VIRUS REASSORTMENT
Abstract
New influenza donor strains for the production of reassortant
influenza A viruses are provided.
Inventors: |
SUPHAPHIPHAT; Pirada;
(Brookline, MA) ; MASON; Peter; (Sommerville,
MA) ; KEINER; Bjoern; (Basel, CH) ; DORMITZER;
Philip; (Weston, MA) ; TRUSHEIM; Heidi; (Apex,
NC) |
Assignee: |
Novartis AG
Basel
CH
|
Family ID: |
47997362 |
Appl. No.: |
13/909013 |
Filed: |
June 3, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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PCT/EP13/54227 |
Mar 2, 2013 |
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13909013 |
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61605922 |
Mar 2, 2012 |
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61685766 |
Mar 23, 2012 |
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Current U.S.
Class: |
424/206.1 ;
435/235.1; 435/239 |
Current CPC
Class: |
A61K 2039/525 20130101;
A61P 31/16 20180101; C12N 2760/16151 20130101; C07K 14/005
20130101; C12N 2760/16134 20130101; C12N 2760/16121 20130101; C12N
2760/16122 20130101; C12N 7/00 20130101; A61K 39/145 20130101; A61K
2039/5252 20130101; C12N 2760/16161 20130101 |
Class at
Publication: |
424/206.1 ;
435/235.1; 435/239 |
International
Class: |
C12N 7/00 20060101
C12N007/00 |
Claims
1. A reassortant influenza A virus comprising two, three, four,
five, or six backbone viral segments from a donor strain, wherein
the donor strain is selected from the group consisting of 105p30
and PR8-X and at least one viral segment derived from a second
influenza strain.
2. The reassortant influenza A virus of claim 1 wherein at least
one backbone viral segment includes a sequence having at least 95%
identity to a sequence selected from the group consisting of SEQ ID
NOs 9-14 or SEQ ID NOs 17-22.
3. The reassortant influenza A virus of claim 1 wherein at least
one backbone viral segment includes the sequence of SEQ ID NO: 17
or SEQ ID NO: 20.
4. The reassortant influenza A virus of claim 2, wherein the virus
comprises backbone segments from two or more donor strains.
5. The reassortant influenza A virus of claim 4, wherein the PB1
and the PB2 viral segments are from the same donor strain.
6. The reassortant influenza A virus of claim 5, wherein the PB1
viral segment has at least 95% identity to SEQ ID NO: 18 and the
PB2 viral segment has at least 95% identity to SEQ ID NO: 19.
7. The reassortant influenza A virus of claim 6, wherein the virus
further comprises a viral segment having at least 95% identity to a
sequence selected from the group consisting of SEQ ID NOs
17-22.
8. The reassortant influenza A virus of claim 3, wherein the virus
comprises the PB2 segment of SEQ ID NO: 19, the PB1 segment of SEQ
ID NO: 18 and the NP segment of SEQ ID NO: 20.
9. The reassortant influenza A virus of claim 1, wherein the virus
has the HA segment from a pandemic influenza strain.
10. A method of preparing a reassortant influenza virus comprising
steps of (i) introducing into a culture host one or more expression
construct(s) which encode(s) the viral segments required to produce
an influenza virus wherein one or more backbone viral segment(s)
is/are from a 105p30 and/or a PR8-X influenza strain and wherein at
least one viral segment is derived from a second influenza strain;
and (ii) culturing the culture host in order to produce reassortant
virus.
11. The method of claim 10 wherein the at least one expression
construct comprises a sequence having at least 90% identity to a
sequence selected from the group consisting of SEQ ID NOs 10-14 and
17-22.
12. The method of claim 10, further comprising the step (iii) of
purifying the reassortant virus obtained in step (ii).
13. The method of claim 10 wherein the at least one viral segment
from the second influenza strain is the HA segment.
14. A method for producing influenza viruses comprising: (a)
infecting a culture host with the reassortant influenza virus of
claim 1; (b) culturing the host from step (a) to produce the virus;
and (c) purifying the virus obtained in step (b).
15. A method of preparing a vaccine, comprising steps of (x)
preparing a virus by the method of claim 14 and (y) preparing
vaccine from the virus.
16. The method of claim 15, wherein (i) the culture host is an
embryonated hen egg, or (ii) the culture host is a mammalian
cell.
17. The method of claim 16, wherein the culture host is (ii) and
the mammalian cell is an MDCK, Vero or PerC6 cell.
18. The method of claim 17, wherein the mammalian cell grows (i)
adherently or (ii) in suspension.
19. The method of claim 18, wherein the mammalian cell is cell line
MDCK 33016 (DSM ACC2219).
20. The method of claim 15, wherein step (y) includes inactivating
the virus.
21. The method of claim 15, wherein the vaccine is a whole virion
vaccine, a split virion vaccine, a surface antigen vaccine or a
virosomal vaccine.
22. The method of claim 15, wherein the vaccine contains less than
10 ng of residual host cell DNA per dose.
23. An influenza vaccine comprising at least one donor polypeptide
encoded by an influenza A backbone viral segment from a donor
strain, wherein the donor strain is 105p30 and a hemagglutinin
polypeptide that is not from influenza strain 105p30.
24. An influenza vaccine comprising two, three, four, five, or six
donor polypeptides encoded by influenza A backbone viral segments
from at least one donor strain, wherein the at least one donor
strain is selected from the group consisting of 105p30 and PR8X and
a hemagglutinin polypeptide that is not from influenza strains
105p30 or PR8X.
25. The influenza vaccine of claim 23, wherein the influenza A
backbone viral segment includes a sequence having at least 95%
identity with a sequence selected from the group consisting of SEQ
ID NOs 9-14 or SEQ ID NOs 17-22.
26. The influenza vaccine of claim 23, wherein the influenza A
backbone viral segment includes the sequence of SEQ ID NO: 17 or
SEQ ID NO: 20.
27. The influenza vaccine of claim 25 wherein the donor
polypeptides are encoded by backbone segments from two or more
donor strains.
28. The influenza vaccine of claim 27, wherein the donor
polypeptides include a PB1 polypeptide and a PB2 polypeptide that
are encoded by influenza A backbone viral segments from the same
donor strain.
29. The influenza vaccine of claim 28, wherein the PB1 polypeptide
is encoded by a backbone viral segment that has at least 95%
identity to SEQ ID NO: 19, and the PB2 polypeptide is encoded by a
backbone viral segment that has at least 95% identity to SEQ ID NO:
19.
30. The influenza vaccine of claim 29, further comprising a donor
polypeptide encoded by a viral segment having at least 95% identity
to a sequence selected from the group consisting of SEQ ID NOs
17-22.
31. The influenza vaccine of claim 23, wherein the vaccine
comprises a PB2 polypeptide encoded by SEQ ID NO: 19, a PB1
polypeptide encoded by SEQ ID NO: 18 and an NP polypeptide encoded
by SEQ ID NO: 20.
32. The influenza vaccine of claim 23, wherein the hemagglutinin is
from a pandemic influenza strain.
Description
[0001] This patent application is a continuation of International
Application No. PCT/EP2013/054227, filed Mar. 2, 2013, which claims
priority from U.S. provisional patent applications 61/605,922,
filed Mar. 2, 2012 and 61/685,766 filed Mar. 23, 2012, the complete
contents of which are incorporated herein by reference.
SUBMISSION OF SEQUENCE LISTING ON ASCII TEXT FILE
[0002] The content of the following submission on ASCII text file
is incorporated herein by reference in its entirety: a computer
readable form (CRF) of the Sequence Listing (file name:
PAT055008_ST25.txt, date recorded: May 22, 2013, size: 161 KB).
TECHNICAL FIELD
[0003] This invention is in the field of influenza A virus
reassortment. Furthermore, it relates to manufacturing vaccines for
protecting against influenza A viruses.
BACKGROUND ART
[0004] The most efficient protection against influenza infection is
vaccination against circulating strains and it is important to
produce influenza viruses for vaccine production as quickly as
possible.
[0005] Wild-type influenza viruses often grow to low titres in eggs
and cell culture. In order to obtain a better-growing virus strain
for vaccine production it is currently common practice to reassort
the circulating vaccine strain with a faster-growing high-yield
donor strain. This can be achieved by co-infecting a culture host
with the circulating influenza strain (the vaccine strain) and the
high-yield donor strain and selecting for reassortant viruses which
contain the hemagglutinin (HA) and neuraminidase (NA) segments from
the vaccine strain and the other viral segments (i.e. those
encoding PB1, PB2, PA, NP, M.sub.1, M.sub.2, NS.sub.1 and NS.sub.2)
from the donor strain. Another approach is to reassort the
influenza viruses by reverse genetics (see, for example references
1 and 2).
[0006] Reference 3 reports that a reassortant influenza virus
containing a PB1 gene segment from A/Texas/1/77, the HA and NA
segments from A/New Caledonia/20/99, a modified PA segment derived
from A/Puerto Rico/8/34 and the remaining viral segments from
A/Puerto Rico/8/34 shows increased growth in cells.
[0007] There are currently only a limited number of donor strains
for reassorting influenza viruses for vaccine manufacture, and the
strain most commonly used is the A/Puerto Rico/8/34 (A/PR/8/34)
strain. However, reassortant influenza viruses comprising A/PR/8/34
backbone segments do not always grow sufficiently well to ensure
efficient vaccine manufacture. Thus, there is a need in the art to
provide further and improved donor strains for influenza virus
reassortment.
SUMMARY OF PREFERRED EMBODIMENTS
[0008] The inventors have now surprisingly discovered that
influenza viruses which comprise backbone segments from two or more
influenza donor strains can grow faster in a culture host compared
with reassortant influenza A viruses which contain all backbone
segments from the same donor strain. In particular, the inventors
have found that influenza viruses which comprise backbone segments
derived from two high-yield donor strains can produce higher yield
reassortants with target vaccine-relevant HA/NA genes than
reassortants made with either of the two original donor
strains.
[0009] In principle, all segments of closely related influenza A
viruses can be specifically reassorted to produce viable viruses,
but only a small fraction of these viruses will be high-growth
reassortants, due to inefficient activities of the resulting viral
components. The inventors have provided backbone combinations that
produce the high yield strains. Reassortant influenza A viruses
comprising backbone segments from two or more influenza donor
strains may contain the PB 1 and the PB2 viral segments from the
same donor strain, in particular the A/New Caledonia/20/1999-like
strain, referred to herein as the 105p30 strain. The PB1 and PB2
viral segments may have at least 95% identity or 100% identity with
the sequence of SEQ ID NO: 2 and/or SEQ ID NO: 3.
[0010] Where the reassortant influenza A virus comprises backbone
segments from two or three donor strains, each donor strain may
provide more than one of the backbone segments of the reassortant
influenza A virus, but one or two of the donor strains can also
provide only a single backbone segment.
[0011] Where the reassortant influenza A virus comprises backbone
segments from two, three, four or five donor strains, one or two of
the donor strains may provide more than one of the backbone
segments of the reassortant influenza A virus. In general the
reassortant influenza A virus cannot comprise more than six
backbone segments. Accordingly, for example, if one of the donor
strains provides five of the viral segments, the reassortant
influenza A virus can only comprise backbone segments from a total
of two different donor strains.
[0012] Where a reassortant influenza A virus comprises the PB1
segment from A/Texas/1/77, it preferably does not comprise the PA,
NP or M segment from A/Puerto Rico/8/34. Where a reassortant
influenza A virus comprises the PA, NP or M segment from A/Puerto
Rico/8/34, it preferably does not comprise the PB1 segment from
A/Texas/1/77. In some embodiments, the invention does not encompass
reassortant influenza A viruses which have the PB1 segment from
A/Texas/1/77 and the PA, NP and M segments from A/Puerto Rico/8/34.
The PB1 segment from A/Texas/1/77 may have the sequence of SEQ ID
NO: 46 and the PA, NP or M segments from A/Puerto Rico/8/34 may
have the sequence of SEQ ID NOs 47, 48 or 49, respectively.
[0013] The inventors have also discovered that variants of known
donor strains can grow to higher viral titres compared to the
original donor strain and can therefore be better donor strains for
reassorting influenza viruses. Examples of such strains are PR8-X
and 105p30.
[0014] Influenza A virus strains of the invention can grow to
higher viral titres in MDCK cells in the same time and under the
same growth conditions compared with A/Puerto Rico/8/34 and/or have
a higher rescue efficiency compared with reassortant influenza
strains that comprise all backbone segments from the same influenza
donor strain. Further provided is a reassortant influenza A virus
comprising at least one backbone viral segment from such an
influenza strain.
[0015] The invention also provides a reassortant influenza A virus
comprising at least one backbone viral segment from a donor strain,
wherein the donor strain is selected from the group consisting of
105p30 and PR8-X. When the at least one backbone viral segment is
the PA segment it may have a sequence having at least 95% or at
least 99% identity with a sequence selected from the group
consisting of SEQ ID NOs: 9 and 17. When the at least one backbone
viral segment is the PB1 segment, it may have a sequence having at
least 95% or at least 99% identity with a sequence selected from
the group consisting of SEQ ID NOs 10 and 18. When the at least one
backbone viral segment is the PB2 segment, it may have a sequence
having at least 95% or at least 99% identity with a sequence
selected from the group consisting of or SEQ ID NOs: 11 and 19.
When the at least one backbone viral segment is the M segment it
may have a sequence having at least 95% or at least 99% identity
with a sequence selected from the group consisting of SEQ ID NOs:
13 and 21. When the at least one backbone viral segment is the NP
segment it may have a sequence having at least 95% or at least 99%
identity with a sequence selected from the group consisting of SEQ
ID NOs: 12 and 20. When the at least one backbone viral segment is
the NS segment it may have a sequence having at least 95% or at
least 99% identity with a sequence selected from the group
consisting of SEQ ID NOs: 14 and 22.
[0016] In embodiments where the reassortant influenza A virus
comprises backbone segments from at least two influenza donor
strains, at least one backbone segment may be derived from a donor
strain selected from the group consisting of 105p30 and PR8-X, as
discussed in the previous paragraph. Preferred reassortant
influenza A viruses comprise 1, 2, 3 or 4 viral segments from the
105p30 donor strain wherein the PA segment may have at least 95%
identity or 100% identity with SEQ ID NO: 17, the NP segment may
have at least 95% identity or 100% identity with SEQ ID NO: 20, the
M segment may have at least 95% identity or 100% identity with SEQ
ID NO: 21, and/or the NS segment may have at least 95% identity or
100% identity with SEQ ID NO: 22. In some embodiments such
influenza A viruses may also comprise at least one backbone viral
segment from the PR8-X donor strain. Where the at least one viral
segment is the PA segment it may have at least 95% identity or 100%
identity with SEQ ID NO: 9. Where the at least one viral segment is
the NP segment it may have at least 95% identity or 100% identity
with SEQ ID NO: 12. Where the at least one viral segment is the M
segment it may have at least 95% identity or 100% identity with SEQ
ID NO: 13. Where the at least one viral segment is the NS segment
it may have at least 95% identity or 100% identity with SEQ ID NO:
9. The inventors have shown that reassortant influenza A viruses
comprising such backbone segments grow well in cell culture. In
general a reassortant influenza virus will contain only one of each
backbone segment. For example, when the influenza virus comprises
the PA segment from 105p30 it will not at the same time comprise
the PA segment of PR8-X.
[0017] In preferred embodiments, the virus comprises viral segments
having at least 95% identity or 100% identity with the sequence of
(a) the PB2 segment of SEQ ID NO: 19, the PB 1 segment of SEQ ID:
NO 18 and the NS segment of SEQ ID NO: 22; or (b) the PB2 segment
of SEQ ID NO: 19, the PB1 segment of SEQ ID NO: 18 and the M
segment of SEQ ID NO: 21; or (c) the PB2 segment of SEQ ID NO: 19,
the PB1 segment of SEQ ID NO: 18 and the NP segment of SEQ ID NO:
20; or (d) the PB2 segment of SEQ ID NO 19, the PB1 segment of SEQ
ID NO 18 and the PA segment of SEQ ID NO 17. These embodiments are
preferred because the inventors have found that such reassortant
influenza A viruses grow particularly well in cell culture.
[0018] The invention provides a method of preparing the reassortant
influenza A viruses of the invention. These methods comprise steps
of (i) introducing into a culture host one or more expression
construct(s) which encode(s) the viral segments required to produce
an influenza A virus wherein the backbone viral segments are from
two or more influenza strains; and (ii) culturing the culture host
in order to produce reassortant virus and optionally (iii)
purifying the virus obtained in step (ii).
[0019] The method may comprise the steps of (i) introducing into a
culture host one or more expression construct(s) which encode(s)
the viral segments required to produce an influenza A virus wherein
the backbone viral segments are from two or more influenza strains
and the PB1 and PB2 segments are from the same donor strain; and
(ii) culturing the culture host in order to produce reassortant
virus and optionally (iii) purifying the virus obtained in step
(ii).
[0020] Also provided is a method of preparing a reassortant
influenza A virus of the invention comprising the steps of (i)
introducing into a culture host one or more expression construct(s)
which encode(s) the viral segments required to produce an influenza
A virus wherein the backbone viral segments are from two or more
influenza strains and the HA and the PB 1 segment are from
different influenza strains which have the same influenza HA
subtype; and (ii) culturing the culture host in order to produce
reassortant virus and optionally (iii) purifying the virus obtained
in step (ii).
[0021] The invention also provides a method of preparing a
reassortant influenza A virus of the invention comprising steps of
(i) introducing into a culture host one or more expression
construct(s) which encode(s) the viral segments required to produce
an influenza A virus wherein one or more backbone viral segment(s)
is/are from a 105p30 and/or a PR8-X influenza strain and wherein at
least one viral segment is derived from a second influenza strain;
and (ii) culturing the culture host in order to produce reassortant
virus and optionally (iii) purifying the virus obtained in step
(ii).
[0022] The methods may further comprise steps of: (iv) infecting a
culture host with the virus obtained in step (ii) or step (iii);
(v) culturing the culture host from step (iv) to produce further
virus; and optionally (vi) purifying the virus obtained in step
(v).
[0023] The invention also provides a method for producing influenza
viruses comprising steps of (a) infecting a culture host with a
reassortant virus of the invention; (b) culturing the host from
step (a) to produce the virus; and optionally (c) purifying the
virus obtained in step (b).
[0024] The invention also provides a method of preparing a vaccine,
comprising steps of (d) preparing a virus by the methods of any one
of the embodiments described above and (e) preparing vaccine from
the virus.
[0025] In a further embodiment, the invention provides influenza
strains PR8-X and 105p30.
[0026] The invention also encompasses variant H1N1 influenza virus
strains in which the M genome segment has lysine in the position
corresponding to amino acid 95 of SEQ ID NO: 33 when aligned to SEQ
ID NO: 33 using a pairwise alignment algorithm. The variant H1N1
influenza virus strains according to the invention may further have
a HA segment which has glycine in the position corresponding to
amino acid 225 of SEQ ID NO: 35 when aligned to SEQ ID NO: 35
and/or has asparagine in the position corresponding to amino acid
231 of SEQ ID NO: 35 when aligned to SEQ ID NO: 35 using a pairwise
alignment algorithm. The variant H1N1 influenza virus strain may
also have a NA segment which has histidine in the position
corresponding to amino acid 70 of SEQ ID NO: 31 when aligned to SEQ
ID NO: 31 using a pairwise alignment algorithm.
[0027] The preferred pairwise alignment algorithm is the
Needleman-Wunsch global alignment algorithm [4], using default
parameters (e.g. with Gap opening penalty=10.0, and with Gap
extension penalty=0.5, using the EBLOSUM62 scoring matrix). This
algorithm is conveniently implemented in the needle tool in the
EMBOSS package [5].
[0028] The invention provides an expression system comprising one
or more expression construct(s) comprising the vRNA encoding
segments of an influenza A virus wherein the expression
construct(s) encode(s) the backbone viral segments from two or more
influenza donor strains. The expression construct(s) may encode the
PB1 and PB2 segments from the same donor strain.
[0029] The invention also provides an expression system comprising
one or more expression construct(s) comprising the vRNA encoding
segments of a 105p30 or PR8-X strain wherein the expression
construct(s) comprise(s) at least one backbone viral segment from
the 105p30 or PR8-X, or strain. The expression construct(s) may
further comprise the vRNAs which encode the PB2, NP, NS, M and PA
segments from PR8-X.
[0030] The invention also provides a host cell comprising the
expression systems of the invention. These host cells can express
an influenza A virus from the expression construct(s) in the
expression system.
[0031] Expression constructs which can be used in the expression
systems of the invention are also provided. For example, the
invention provides an expression construct which encodes the
backbone segments of the reassortant influenza strains according to
the invention on the same construct.
Donor Strains
[0032] Influenza donor strains are strains which typically provide
the backbone segments in a reassortant influenza virus, even though
they may sometimes also provide the HA or NA segment, but not both,
of the virus. Usually, however, both the HA and the NA segment in a
reassortant influenza virus will be from the vaccine strain.
[0033] The inventors have surprisingly discovered that reassortant
influenza A viruses comprising backbone segments from two or more
influenza donor strains can grow to higher titres in cell culture
compared with reassortant influenza viruses which contain all
backbone segments from the same donor strain. The inventors have
shown that this effect is due to the presence of backbone segments
from two donor strains and does not require the presence of viral
segments with specific mutations. Particularly good results are
achieved, however, with influenza A strains in which the M genome
segment has lysine in the position corresponding to amino acid 95
of SEQ ID NO: 33 when aligned to SEQ ID NO: 33.
[0034] Reassortant influenza A viruses comprising the PB1 and PB2
segments from the same influenza strain (for example 105p30) are
also advantageous because they showed a better rescue efficiency
compared with influenza viruses in which the PB1 and PB2 segments
are from different viruses. The PB1 and PB2 segments of 105p30 have
the sequence of SEQ ID NOs 18 and 19, respectively.
[0035] The inventors have also shown that some influenza virus
strains can grow to higher viral titres in MDCK cells in the same
time and under the same growth conditions compared with A/Puerto
Rico/8/34.
[0036] Variants of influenza donor strains which are derived from
the donor strains of the invention or other known donor strains
such A/PR/8/34 (wt PR8) can also be useful as donor strains. These
donor strains can grow to higher viral titres (in the same time and
under the same growth conditions) compared to the donor strain from
which they are derived. For example, the inventors have
surprisingly discovered that passaging the A/PR/8/34 influenza
strain several times in cell culture results in a virus strain
(PR8-X) which grows to much higher viral titres compared to the
original A/PR8/34 strain. Likewise, the inventors have found that
passaging the A/New Caledonia/20/1999 strain several times in cells
results in a strain (105p30) which grows to even higher viral
titres compared to the unpassaged A/New Caledonia/20/1999 strain in
the same time and under the same growth conditions. Donor strain
variants of the present invention will typically achieve viral
titres which are at least 10%, at least 20%, at least 50%, at least
100%, at least 200%, at least 500% or at least 1000% higher under
the same growth conditions and for the same time (for example 12
hours, 24 hours, 48 hours or 72 hours) compared to the viral titres
obtained with the donor strain from which the variant was
derived.
[0037] The segments of PR8-X have the sequences of SEQ ID NO: 11
(PB2), SEQ ID NO: 10 (PB1), SEQ ID NO: 9 (PA), SEQ ID NO: 12 (NP),
SEQ ID NO: 13 (M), SEQ ID NO: 14 (NS), SEQ ID NO: 15 (HA) or SEQ ID
NO: 16 (NA).
[0038] The segments of 105p30 have the sequences of SEQ ID NO: 19
(PB2), SEQ ID NO: 18 (PB1), SEQ ID NO: 17 (PA), SEQ ID NO: 20 (NP),
SEQ ID NO: 21 (M), SEQ ID NO: 22 (NS), SEQ ID NO: 23 (HA) or SEQ ID
NO: 24 (NA).
[0039] Influenza strains which contain one, two, three, four five,
six or seven of the segments of the 105p30 or PR8-X strains can
also be used as donor strains.
[0040] The invention can be practised with donor strains having a
viral segment that has at least about 70%, at least about 75%, at
least about 80%, at least about 85%, at least about 90%, at least
about 95% or at least about 99% identity to a sequence of SEQ ID
NOs 11-14 or 18-22. For example, due to the degeneracy of the
genetic code, it is possible to have the same polypeptide encoded
by several nucleic acids with different sequences. Thus, the
invention may be practised with viral segments that encode the same
polypeptides as the sequences of SEQ ID NOs 11-14 or 18-22. For
example, the nucleic acid sequences of SEQ ID NOs: 3 and 28 have
only 73% identity even though they encode the same viral
protein.
[0041] The invention may also be practised with viral segments that
encode polypeptides that have at least 80%, at least 85%, at least
90%, at least 95% or at least 99% identity to the polypeptide
sequences encoded by SEQ ID NOs 11-14 or 18-22.
[0042] Variations in the DNA and the amino acid sequence may also
stem from spontaneous mutations which can occur during passaging of
the viruses. Such variant influenza strains can also be used in the
invention.
Reassortant Viruses
[0043] The invention provides reassortant influenza viruses which
comprise backbone segments from two or more influenza donor
strains. The PB 1 and PB2 segments may be from the same donor
strain.
[0044] The invention also provides reassortant influenza viruses
comprising at least one backbone viral segment from an influenza
virus strain that can grow to higher viral titres in MDCK cells in
the same time and under the same growth conditions compared with
A/Puerto Rico/8/34.
[0045] The invention provides reassortant influenza viruses
comprising at least one backbone viral segment from the donor
strains of the invention, e.g. a PR8-X or 105p30 strain. The
reassortant influenza viruses of the invention can be reassortants
between two, three or more different influenza strains provided
that at least one viral segment is derived from a donor strain of
the invention.
[0046] Influenza viruses are segmented negative strand RNA viruses.
Influenza A and B viruses have eight segments (NP, M, NS, PA, PB1,
HA and NA) whereas influenza C virus has seven. The reassortant
viruses of the invention contain the backbone segments from two or
more donor strains, or at least one (i.e. one, two, three, four,
five or six) backbone viral segment from the donor strains of the
invention. The backbone viral segments are those which do not
encode HA or NA. Thus, backbone segments will typically encode the
PB1, PB2, PA, NP, M.sub.1, M.sub.2, NS.sub.1 and NS.sub.2
polypeptides of the influenza virus. The reassortant viruses will
not typically contain the segments encoding HA and NA from the
donor strains even though reassortant viruses which comprise either
the HA or the NA but not both from the donor strains of the
invention are also envisioned.
[0047] When the reassortant viruses of the invention are
reassortants comprising the backbone segments from a single donor
strain, the reassortant viruses will generally include segments
from the donor strain and the vaccine strain in a ratio of 1:7,
2:6, 3:5, 4:4, 5:3, 6:2 or 7:1. Having a majority of segments from
the donor strain, in particular a ratio of 6:2, is typical. When
the reassortant viruses comprise backbone segments from two donor
strains, the reassortant virus will generally include segments from
the first donor strain, the seconds donor strain and the vaccine
strain in a ratio of 1:1:6, 1:2:5, 1:3:4, 1:4:3, 1:5:2, 1:6:1,
2:1:5, 2:2:4, 2:3:3, 2:4:2, 2:5:1, 3:1:2, 3:2:1, 4:1:3, 4:2:2,
4:3:1, 5:1:2, 5:2:1 or 6:1:1.
[0048] Preferably, the reassortant viruses do not contain the HA
segment of the donor strain as this encodes the main vaccine
antigens of the influenza virus and should therefore come from the
vaccine strain. The reassortant viruses of the invention therefore
preferably have at least the HA segment and typically the HA and NA
segments from the vaccine strain.
[0049] The invention also encompasses reassortant viruses which
contain viral segments from more than one, for example two or three
different, donor strain(s) wherein at least one viral segment,
preferably not HA, is derived from the PR8-X or 105p30 influenza
strains. Such reassortant influenza viruses will typically contain
the HA and/or NA segment from a vaccine strain. Where the
reassortants contain viral segments from more than one influenza
donor strain, the further donor strain(s) can be any donor strain
including the donor strains of the invention. For example, some of
the viral segments may be derived from the A/PR/8/34 or AA/6/60
(A/Ann Arbor/6/60) influenza strains. Reassortants containing viral
segments from the AA/6/60 strain may be advantageous, for example,
where the reassortant virus is to be used in a live attenuated
influenza vaccine.
[0050] The invention also encompasses reassortants which comprise
viral segments from more than one vaccine strain provided that the
reassortant comprises a backbone according to the present
invention. For example, the reassortant influenza viruses may
comprise the HA segment from one donor strain and the NA segment
from a different donor strain.
[0051] The reassortant viruses of the invention can grow to higher
viral titres than the wild-type vaccine strain from which some of
the viral segment(s) of the reassortant virus are derived in the
same time (for example 12 hours, 24 hours, 48 hours or 72 hours)
and under the same growth conditions. The viral titre can be
determined by standard methods known to those of skill in the art.
The reassortant viruses of the invention can achieve a viral titre
which is at least 10% higher, at least 20% higher, at least 50%
higher, at least 100% higher, at least 200% higher, at least 500%
higher, or at least 1000% higher than the viral titre of the wild
type vaccine strain in the same time frame and under the same
conditions.
[0052] The invention is suitable for reassorting pandemic as well
as inter-pandemic (seasonal) influenza vaccine strains. The
reassortant influenza strains may contain the influenza A virus HA
subtypes H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13,
H14, H15 or H16. They may contain the influenza A virus NA subtypes
N1, N2, N3, N4, N5, N6, N7, N8 or N9. Where the vaccine strain used
in the reassortant influenza viruses of the invention is a seasonal
influenza strain, the vaccine strain may have a H1 or H3 subtype.
In one aspect of the invention the vaccine strain is a H1N1 or
H.sub.3N.sub.2 strain.
[0053] The vaccine strains for use in the invention may also be
pandemic strains or potentially pandemic strains. The
characteristics of an influenza strain that give it the potential
to cause a pandemic outbreak are: (a) it contains a new
hemagglutinin compared to the hemagglutinins in
currently-circulating human strains, i.e. one that has not been
evident in the human population for over a decade (e.g. H2), or has
not previously been seen at all in the human population (e.g. H5,
H6 or H9, that have generally been found only in bird populations),
such that the human population will be immunologically naive to the
strain's hemagglutinin; (b) it is capable of being transmitted
horizontally in the human population; and (c) it is pathogenic to
humans. A vaccine strain with H5 hemagglutinin type is preferred
where the reassortant virus is used in vaccines for immunizing
against pandemic influenza, such as a H5N1 strain. Other possible
strains include H5N3, H9N2, H2N2, H7N1 and H7N7, and any other
emerging potentially pandemic strains. The invention is
particularly suitable for producing reassortant viruses for use in
vaccine for protecting against potential pandemic virus strains
that can or have spread from a non-human animal population to
humans, for example a swine-origin H1N1 influenza strain.
[0054] The reassortant influenza strain of the invention may
comprise the HA segment and/or the NA segment from an
A/California/4/09 strain. Thus, for instance, the HA gene segment
may encode a H1 hemagglutinin which is more closely related to SEQ
ID NO: 32 than to SEQ ID NO: 25 (i.e. has a higher degree sequence
identity when compared to SEQ ID NO: 32 than to SEQ ID NO: 25 using
the same algorithm and parameters). SEQ ID NOs: 32 and 25 are 80%
identical. Similarly, the NA gene may encode a N1 neuraminidase
which is more closely related to SEQ ID NO: 27 than to SEQ ID NO:
26. SEQ ID NOs: 27 and 26 are 82% identical.
[0055] Strains which can be used as vaccine strains include strains
which are resistant to antiviral therapy (e.g. resistant to
oseltamivir [6] and/or zanamivir), including resistant pandemic
strains [7].
[0056] The choice of donor strain for use in the methods of the
invention can depend on the vaccine strain which is to be
reassorted. As reassortants between evolutionary distant strains
might not replicate well in cell culture, it is possible that the
donor strain and the vaccine strain have the same HA and/or NA
subtype. In other embodiments, however, the vaccine strain and the
donor strain can have different HA and/or NA subtypes, and this
arrangement can facilitate selection for reassortant viruses that
contain the HA and/or NA segment from the vaccine strain.
Therefore, although the 105p30 and PR8-X strains contain the H1
influenza subtype these donor strains can be used for vaccine
strains which do not contain the H1 influenza subtype.
[0057] Reassortants of the donor strains of the invention wherein
the HA and/or NA segment has been changed to another subtype can
also be used. The H1 influenza subtype of the 105p30 or PR8-X
strain may be changed, for example, to a H3 or H5 subtype.
[0058] Thus, the invention encompasses an influenza A virus which
comprises one, two, three, four, five, six or seven viral segments
from the 105p30 or PR8-X strains of the invention and a HA segment
which is not of the H1 subtype. The reassortant donor strains may
further comprise an NA segment which is not of the N1 subtype.
Suitable techniques for reassorting the donor strains will be
evident to those of skill in the art.
[0059] The invention also encompasses reassortant donor strains
which comprise at least one, at least two, at least three, at least
four, at least five, at least six or at least seven viral segments
from the 105p30 or PR8-X strains of the invention and a H1 HA
segment which is derived from a different influenza strain.
[0060] Reassortant viruses which contain an NS segment that does
not encode a functional NS protein are also within the scope of the
present invention. NS 1 knockout mutants are described in reference
8. These NS1-mutant virus strains are particularly suitable for
preparing live attenuated influenza vaccines.
[0061] The `second influenza strain` used in the methods of the
invention is different to the donor strain which is used.
Reverse Genetics
[0062] The invention is particularly suitable for producing
reassortant influenza virus strains through reverse genetics
techniques. In these techniques, the viruses are produced in
culture hosts using an expression system.
[0063] In one aspect, the expression system may encode the PB1 and
PB2 segments from the same donor strain. In this aspect of the
invention, the system may encode at least one (i.e. one, two three
or four) of the segments NP, M, NS and/or PA from another influenza
donor strain.
[0064] In another aspect, the system may encode 1 or more (e.g. 1,
2, 3, 4, 5 or 6) genome segments from the PR8-X strain, but usually
this/these will not include the PR8-X HA segment and usually will
not include the PR8-X NA segment. Thus the system may encode at
least one of segments NP, M, NS, PA, PB1 and/or PB2 (possibly all
six) from PR8-X.
[0065] The system may encode 1 or more (e.g. 1, 2, 3, 4, 5 or 6)
genome segments from the 105p30 strain, but usually this/these will
not include the 105p30 HA segment and usually will not include the
105p30 NA segment. Thus the system may encode at least one of
segments NP, M, NS, PA, PB1 and/or PB2 (possibly all six) from
105p30.
[0066] Reverse genetics for influenza A and B viruses can be
practised with 12 plasmids to express the four proteins required to
initiate replication and transcription (PB 1, PB2, PA and
nucleoprotein) and all eight viral genome segments. To reduce the
number of constructs, however, a plurality of RNA polymerase I
transcription cassettes (for viral RNA synthesis) can be included
on a single plasmid (e.g. sequences encoding 1, 2, 3, 4, 5, 6, 7 or
all 8 influenza vRNA segments), and a plurality of protein-coding
regions with RNA polymerase II promoters on another plasmid (e.g.
sequences encoding 1, 2, 3, 4, 5, 6, 7 or 8 influenza mRNA
transcripts) [9]. It is also possible to include one or more
influenza vRNA segments under control of a pol I promoter and one
or more influenza protein coding regions under control of another
promoter, in particular a pol II promoter, on the same plasmid.
This is preferably done by using bi-directional plasmids.
[0067] Preferred aspects of the reference 9 method involve: (a) PB
1, PB2 and PA mRNA-encoding regions on a single expression
construct; and (b) all 8 vRNA encoding segments on a single
expression construct. Including the neuraminidase (NA) and
hemagglutinin (HA) segments on one expression construct and the six
other viral segments on another expression construct is
particularly preferred as newly emerging influenza virus strains
usually have mutations in the NA and/or HA segments. Therefore, the
advantage of having the HA and/or NA segments on a separate
expression construct is that only the vector comprising the HA and
NA sequence needs to be replaced. Thus, in one aspect of the
invention the NA and/or HA segments of the vaccine strain may be
included on one expression construct and the vRNA encoding segments
from the donor strain(s) of the invention, excluding the HA and/or
NA segment(s), are included on a different expression construct.
The invention thus provides an expression construct comprising one,
two, three, four, five or six vRNA encoding backbone viral segments
of a donor strain of the invention. The expression construct may
not comprise HA and/or NA viral segments that produce a functional
HA and/or NA protein.
[0068] Known reverse genetics systems involve expressing DNA
molecules which encode desired viral RNA (vRNA) molecules from pol
I promoters, bacterial RNA polymerase promoters, bacteriophage
polymerase promoters, etc. As influenza viruses require the
presence of viral polymerase to complete the life cycle, systems
may also provide these proteins e.g. the system further comprises
DNA molecules that encode viral polymerase proteins such that
expression of both types of DNA leads to assembly of a complete
infectious virus. It is also possible to supply the viral
polymerase as a protein.
[0069] Where reverse genetics is used for the expression of
influenza vRNA, it will be evident to the person skilled in the art
that precise spacing of the sequence elements with reference to
each other is important for the polymerase to initiate replication.
It is therefore important that the DNA molecule encoding the viral
RNA is positioned correctly between the pol I promoter and the
termination sequence, but this positioning is well within the
capabilities of those who work with reverse genetics systems.
[0070] In order to produce a recombinant virus, a cell must express
all segments of the viral genome which are necessary to assemble a
virion. DNA cloned into the expression constructs of the present
invention preferably provides all of the viral RNA and proteins,
but it is also possible to use a helper virus to provide some of
the RNA and proteins, although systems which do not use a helper
virus are preferred. As the influenza virus is a segmented virus,
the viral genome will usually be expressed using more than one
expression construct in the methods of the invention. It is also
envisioned, however, to combine one or more segments or even all
segments of the viral genome on a single expression construct.
[0071] In some embodiments an expression construct will also be
included which leads to expression of an accessory protein in the
host cell. For instance, it can be advantageous to express a
non-viral serine protease (e.g. trypsin) as part of a reverse
genetics system.
Expression Constructs
[0072] Expression constructs used in the expression systems of the
invention may be uni-directional or bi-directional expression
constructs. Where more than one transgene is used in the methods
(whether on the same or different expression constructs) it is
possible to use uni-directional and/or bi-directional
expression.
[0073] As influenza viruses require a protein for infectivity, it
is generally preferred to use bi-directional expression constructs
as this reduces the total number of expression constructs required
by the host cell. Thus, the method of the invention may utilise at
least one bi-directional expression construct wherein a gene or
cDNA is located between an upstream pol II promoter and a
downstream non-endogenous pol I promoter. Transcription of the gene
or cDNA from the pol II promoter produces capped positive-sense
viral mRNA which can be translated into a protein, while
transcription from the non-endogenous pol I promoter produces
negative-sense vRNA. The bi-directional expression construct may be
a bi-directional expression vector.
[0074] Bi-directional expression constructs contain at least two
promoters which drive expression in different directions (i.e. both
5' to 3' and 3' to 5') from the same construct. The two promoters
can be operably linked to different strands of the same double
stranded DNA. Preferably, one of the promoters is a pol I promoter
and at least one of the other promoters is a pol II promoter. This
is useful as the pol I promoter can be used to express uncapped
vRNAs while the pol II promoter can be used to transcribe mRNAs
which can subsequently be translated into proteins, thus allowing
simultaneous expression of RNA and protein from the same construct.
Where more than one expression construct is used within an
expression system, the promoters may be a mixture of endogenous and
non-endogenous promoters.
[0075] The pol I and pol II promoters used in the expression
constructs may be endogenous to an organism from the same taxonomic
order from which the host cell is derived. Alternatively, the
promoters can be derived from an organism in a different taxonomic
order than the host cell. The term "order" refers to conventional
taxonomic ranking, and examples of orders are primates, rodentia,
carnivora, marsupialia, cetacean, etc. Humans and chimpanzees are
in the same taxonomic order (primates), but humans and dogs are in
different orders (primates vs. carnivora). For example, the human
pol I promoter can be used to express viral segments in canine
cells (e.g. MDCK cells).
[0076] The expression construct will typically include an RNA
transcription termination sequence. The termination sequence may be
an endogenous termination sequence or a termination sequence which
is not endogenous to the host cell. Suitable termination sequences
will be evident to those of skill in the art and include, but are
not limited to, RNA polymerase I transcription termination
sequence, RNA polymerase II transcription termination sequence, and
ribozymes. Furthermore, the expression constructs may contain one
or more polyadenylation signals for mRNAs, particularly at the end
of a gene whose expression is controlled by a pol II promoter.
[0077] An expression system may contain at least two, at least
three, at least four, at least five, at least six, at least seven,
at least eight, at least nine, at least ten, at least eleven or at
least twelve expression constructs.
[0078] An expression construct may be a vector, such as a plasmid
or other episomal construct. Such vectors will typically comprise
at least one bacterial and/or eukaryotic origin of replication.
Furthermore, the vector may comprise a selectable marker which
allows for selection in prokaryotic and/or eukaryotic cells.
Examples of such selectable markers are genes conferring resistance
to antibiotics, such as ampicillin or kanamycin. The vector may
further comprise one or more multiple cloning sites to facilitate
cloning of a DNA sequence.
[0079] As an alternative, an expression construct may be a linear
expression construct. Such linear expression constructs will
typically not contain any amplification and/or selection sequences.
However, linear constructs comprising such amplification and/or
selection sequences are also within the scope of the present
invention. Reference 10 describes a linear expression construct
which describes individual linear expression constructs for each
viral segment. It is also possible to include more than one, for
example two, three four, five or six viral segments on the same
linear expression construct. Such a system has been described, for
example, in reference 11.
[0080] Expression constructs can be generated using methods known
in the art. Such methods were described, for example, in reference
12. Where the expression construct is a linear expression
construct, it is possible to linearise it before introduction into
the host cell utilising a single restriction enzyme site.
Alternatively, it is possible to excise the expression construct
from a vector using at least two restriction enzyme sites.
Furthermore, it is also possible to obtain a linear expression
construct by amplifying it using a nucleic acid amplification
technique (e.g. by PCR).
[0081] The expression constructs used in the systems of the
invention may be non-bacterial expression constructs. This means
that the construct can drive expression in a eukaryotic cell of
viral RNA segments encoded therein, but it does not include
components which would be required for propagation of the construct
in bacteria. Thus the construct will not include a bacterial origin
of replication (ori), and usually will not include a bacterial
selection marker (e.g. an antibiotic resistance marker). Such
expression constructs are described in reference 13 which is
incorporated by reference.
[0082] The expression constructs may be prepared by chemical
synthesis. The expression constructs may either be prepared
entirely by chemical synthesis or in part. Suitable methods for
preparing expression constructs by chemical synthesis are
described, for example, in reference 13 which is incorporated by
reference.
[0083] The expression constructs of the invention can be introduced
into host cells using any technique known to those of skill in the
art. For example, expression constructs of the invention can be
introduced into host cells by employing electroporation,
DEAE-dextran, calcium phosphate precipitation, liposomes,
microinjection, or microparticle-bombardment.
Cells
[0084] The culture host for use in the present invention can be any
eukaryotic cell that can produce the virus of interest. The
invention will typically use a cell line although, for example,
primary cells may be used as an alternative. The cell will
typically be mammalian. Suitable mammalian cells include, but are
not limited to, hamster, cattle, primate (including humans and
monkeys) and dog cells. Various cell types may be used, such as
kidney cells, fibroblasts, retinal cells, lung cells, etc. Examples
of suitable hamster cells are the cell lines having the names BHK21
or HKCC. Suitable monkey cells are e.g. African green monkey cells,
such as kidney cells as in the Vero cell line [14-15]. Suitable dog
cells are e.g. kidney cells, as in the CLDK and MDCK cell
lines.
[0085] Further suitable cells include, but are not limited to: CHO;
293T; BHK; MRC 5; PER.C6 [16]; FRhL2; WI-38; etc. Suitable cells
are widely available e.g. from the American Type Cell Culture
(ATCC) collection [17], from the Coriell Cell Repositories [18], or
from the European Collection of Cell Cultures (ECACC). For example,
the ATCC supplies various different Vero cells under catalogue
numbers CCL 81, CCL 81.2, CRL 1586 and CRL-1587, and it supplies
MDCK cells under catalogue number CCL 34. PERC6 is available from
the ECACC under deposit number 96022940.
[0086] Preferred cells for use in the invention are MDCK cells
[19-20], derived from Madill Darby canine kidney. The original MDCK
cells are available from the ATCC as CCL 34. It is preferred that
derivatives of MDCK cells are used. Such derivatives were
described, for instance, in reference 19 which discloses MDCK cells
that were adapted for growth in suspension culture (`MDCK 33016` or
`33016-PF`, deposited as DSM ACC 2219; see also ref. 19).
Furthermore, reference 21 discloses MDCK-derived cells that grow in
suspension in serum free culture (`B-702`, deposited as FERM
BP-7449). In some embodiments, the MDCK cell line used may be
tumorigenic. It is also envisioned to use non-tumorigenic MDCK
cells. For example, reference 22 discloses non tumorigenic MDCK
cells, including `MDCK-S` (ATCC PTA-6500), `MDCK-SF101` (ATCC
PTA-6501), `MDCK-SF102` (ATCC PTA-6502) and `MDCK-SF103` (ATCC
PTA-6503). Reference 23 discloses MDCK cells with high
susceptibility to infection, including `MDCK.5F1` cells (ATCC CRL
12042).
[0087] It is possible to use a mixture of more than one cell type
to practise the methods of the present invention. However, it is
preferred that the methods of the invention are practised with a
single cell type e.g. with monoclonal cells. Preferably, the cells
used in the methods of the present invention are from a single cell
line. Furthermore, the same cell line may be used for reassorting
the virus and for any subsequent propagation of the virus.
[0088] Preferably, the cells are cultured in the absence of serum,
to avoid a common source of contaminants. Various serum-free media
for eukaryotic cell culture are known to the person skilled in the
art (e.g. Iscove's medium, ultra CHO medium (BioWhittaker), EX-CELL
(JRH Biosciences)). Furthermore, protein-free media may be used
(e.g. PF-CHO (JRH Biosciences)). Otherwise, the cells for
replication can also be cultured in the customary serum-containing
media (e.g. MEM or DMEM medium with 0.5% to 10% of fetal calf
serum).
[0089] The cells may be in adherent culture or in suspension.
Conventional Reassortment
[0090] Traditionally, influenza viruses are reassorted by
co-infecting a culture host, usually eggs, with a donor strain and
a vaccine strain. Reassortant viruses are selected by adding
antibodies with specificity for the HA and/or NA proteins of the
donor strain in order to select for reassortant viruses that
contain the vaccine strain's HA and/or NA proteins. Over several
passages of this treatment one can select for fast growing
reassortant viruses containing the vaccine strain's HA and/or NA
segments.
[0091] The invention is suitable for use in these methods. It can
be easier to use vaccine strains with a different HA and/or NA
subtype compared to the donor strain(s) as this facilitates
selection for reassortant viruses. It is also possible, however, to
use vaccine strains with the same HA and/or NA subtype as the donor
strain(s) and in some aspects of the invention this preferred. In
this case, antibodies with preferential specificity for the HA
and/or NA proteins of the donor strain(s) should be available.
Virus Preparation
[0092] In one embodiment, the invention provides a method for
producing influenza viruses comprising steps of (a) infecting a
culture host with a reassortant virus of the invention; (b)
culturing the host from step (a) to produce the virus; and
optionally (c) purifying the virus obtained in step (b).
[0093] The culture host may be cells or embryonated hen eggs. Where
cells are used as a culture host in this aspect of the invention,
it is known that cell culture conditions (e.g. temperature, cell
density, pH value, etc.) are variable over a wide range subject to
the cell line and the virus employed and can be adapted to the
requirements of the application. The following information
therefore merely represents guidelines.
[0094] As mentioned above, cells are preferably cultured in
serum-free or protein-free media.
[0095] Multiplication of the cells can be conducted in accordance
with methods known to those of skill in the art. For example, the
cells can be cultivated in a perfusion system using ordinary
support methods like centrifugation or filtration. Moreover, the
cells can be multiplied according to the invention in a fed-batch
system before infection. In the context of the present invention, a
culture system is referred to as a fed-batch system in which the
cells are initially cultured in a batch system and depletion of
nutrients (or part of the nutrients) in the medium is compensated
by controlled feeding of concentrated nutrients. It can be
advantageous to adjust the pH value of the medium during
multiplication of cells before infection to a value between pH 6.6
and pH 7.8 and especially between a value between pH 7.2 and pH
7.3. Culturing of cells preferably occurs at a temperature between
30 and 40.degree. C. When culturing the infected cells (step ii),
the cells are preferably cultured at a temperature of between
30.degree. C. and 36.degree. C. or between 32.degree. C. and
34.degree. C. or at 33.degree. C. This is particularly preferred,
as it has been shown that incubation of infected cells in this
temperature range results in production of a virus that results in
improved efficacy when formulated into a vaccine [24].
[0096] Oxygen partial pressure can be adjusted during culturing
before infection preferably at a value between 25% and 95% and
especially at a value between 35% and 60%. The values for the
oxygen partial pressure stated in the context of the invention are
based on saturation of air. Infection of cells occurs at a cell
density of preferably about 8-25.times.10.sup.5 cells/mL in the
batch system or preferably about 5-20.times.10.sup.6 cells/mL in
the perfusion system. The cells can be infected with a viral dose
(MOI value, "multiplicity of infection"; corresponds to the number
of virus units per cell at the time of infection) between 10.sup.-8
and 10, preferably between 0.0001 and 0.5.
[0097] Virus may be grown on cells in adherent culture or in
suspension. Microcarrier cultures can be used. In some embodiments,
the cells may thus be adapted for growth in suspension.
[0098] The methods according to the invention also include
harvesting and isolation of viruses or the proteins generated by
them. During isolation of viruses or proteins, the cells are
separated from the culture medium by standard methods like
separation, filtration or ultrafiltration. The viruses or the
proteins are then concentrated according to methods sufficiently
known to those skilled in the art, like gradient centrifugation,
filtration, precipitation, chromatography, etc., and then purified.
It is also preferred according to the invention that the viruses
are inactivated during or after purification. Virus inactivation
can occur, for example, by .beta.-propiolactone or formaldehyde at
any point within the purification process.
[0099] The culture host may be eggs. The current standard method
for influenza virus growth for vaccines uses embryonated SPF hen
eggs, with virus being purified from the egg contents (allantoic
fluid). It is also possible to passage a virus through eggs and
subsequently propagate it in cell culture and vice versa.
Vaccine
[0100] The invention utilises virus produced according to the
method to produce vaccines.
[0101] Vaccines (particularly for influenza virus) are generally
based either on live virus or on inactivated virus. Inactivated
vaccines may be based on whole virions, `split` virions, or on
purified surface antigens. Antigens can also be presented in the
form of virosomes. The invention can be used for manufacturing any
of these types of vaccine.
[0102] Where an inactivated virus is used, the vaccine may comprise
whole virion, split virion, or purified surface antigens (for
influenza, including hemagglutinin and, usually, also including
neuraminidase). Chemical means for inactivating a virus include
treatment with an effective amount of one or more of the following
agents: detergents, formaldehyde, .beta.-propiolactone, methylene
blue, psoralen, carboxyfullerene (C60), binary ethylamine, acetyl
ethyleneimine, or combinations thereof. Non-chemical methods of
viral inactivation are known in the art, such as for example UV
light or gamma irradiation.
[0103] Virions can be harvested from virus-containing fluids, e.g.
allantoic fluid or cell culture supernatant, by various methods.
For example, a purification process may involve zonal
centrifugation using a linear sucrose gradient solution that
includes detergent to disrupt the virions. Antigens may then be
purified, after optional dilution, by diafiltration.
[0104] Split virions are obtained by treating purified virions with
detergents (e.g. ethyl ether, polysorbate 80, deoxycholate,
tri-N-butyl phosphate, Triton X-100, Triton N101,
cetyltrimethylammonium bromide, Tergitol NP9, etc.) to produce
subvirion preparations, including the `Tween-ether` splitting
process. Methods of splitting influenza viruses, for example are
well known in the art e.g. see refs. 25-26, etc. Splitting of the
virus is typically carried out by disrupting or fragmenting whole
virus, whether infectious or non-infectious with a disrupting
concentration of a splitting agent. The disruption results in a
full or partial solubilisation of the virus proteins, altering the
integrity of the virus. Preferred splitting agents are non-ionic
and ionic (e.g. cationic) surfactants e.g. alkylglycosides,
alkylthioglycosides, acyl sugars, sulphobetaines, betains,
polyoxyethylenealkylethers, N,N-dialkyl-Glucamides, Hecameg,
alkylphenoxy-polyethoxyethanols, NP9, quaternary ammonium
compounds, sarcosyl, CTABs (cetyl trimethyl ammonium bromides),
tri-N-butyl phosphate, Cetavlon, myristyltrimethylammonium salts,
lipofectin, lipofectamine, and DOT-MA, the octyl- or nonylphenoxy
polyoxyethanols (e.g. the Triton surfactants, such as Triton X-100
or Triton N101), polyoxyethylene sorbitan esters (the Tween
surfactants), polyoxyethylene ethers, polyoxyethlene esters, etc.
One useful splitting procedure uses the consecutive effects of
sodium deoxycholate and formaldehyde, and splitting can take place
during initial virion purification (e.g. in a sucrose density
gradient solution). Thus a splitting process can involve
clarification of the virion-containing material (to remove
non-virion material), concentration of the harvested virions (e.g.
using an adsorption method, such as CaHPO.sub.4 adsorption),
separation of whole virions from non-virion material, splitting of
virions using a splitting agent in a density gradient
centrifugation step (e.g. using a sucrose gradient that contains a
splitting agent such as sodium deoxycholate), and then filtration
(e.g. ultrafiltration) to remove undesired materials. Split virions
can usefully be resuspended in sodium phosphate-buffered isotonic
sodium chloride solution. Examples of split influenza vaccines are
the BEGRIVAC.TM., FLUARIX.TM., FLUZONE.TM. and FLUSHIELD.TM.
products.
[0105] Purified influenza virus surface antigen vaccines comprise
the surface antigens hemagglutinin and, typically, also
neuraminidase. Processes for preparing these proteins in purified
form are well known in the art. The FLUVIRIN.TM., AGRIPPAL.TM. and
INFLUVAC.TM. products are influenza subunit vaccines.
[0106] Another form of inactivated antigen is the virosome [27]
(nucleic acid free viral-like liposomal particles). Virosomes can
be prepared by solubilization of virus with a detergent followed by
removal of the nucleocapsid and reconstitution of the membrane
containing the viral glycoproteins. An alternative method for
preparing virosomes involves adding viral membrane glycoproteins to
excess amounts of phospholipids, to give liposomes with viral
proteins in their membrane.
[0107] The methods of the invention may also be used to produce
live vaccines. Such vaccines are usually prepared by purifying
virions from virion-containing fluids. For example, the fluids may
be clarified by centrifugation, and stabilized with buffer (e.g.
containing sucrose, potassium phosphate, and monosodium glutamate).
Various forms of influenza virus vaccine are currently available
(e.g. see chapters 17 & 18 of reference 28). Live virus
vaccines include MedImmune's FLUMIST.TM. product (trivalent live
virus vaccine).
[0108] The virus may be attenuated. The virus may be
temperature-sensitive. The virus may be cold-adapted. These three
features are particularly useful when using live virus as an
antigen.
[0109] HA is the main immunogen in current inactivated influenza
vaccines, and vaccine doses are standardised by reference to HA
levels, typically measured by SRID. Existing vaccines typically
contain about 15 .mu.g of HA per strain, although lower doses can
be used e.g. for children, or in pandemic situations, or when using
an adjuvant. Fractional doses such as 1/2 (i.e. 7.5 .mu.g HA per
strain), 1/4 and 1/8 have been used, as have higher doses (e.g.
3.times. or 9.times. doses [29,30]). Thus vaccines may include
between 0.1 and 150 .mu.g of HA per influenza strain, preferably
between 0.1 and 50 .mu.g e.g. 0.1-20 .mu.g, 0.1-15 .mu.g, 0.1-10
.mu.g, 0.5-5 .mu.g, etc. Particular doses include e.g. about 45,
about 30, about 15, about 10, about 7.5, about 5, about 3.8, about
3.75, about 1.9, about 1.5, etc. per strain.
[0110] For live vaccines, dosing is measured by median tissue
culture infectious dose (TCID.sub.50) rather than HA content, and a
TCID.sub.50 of between 10.sup.6 and 10.sup.8 (preferably between
10.sup.6.5-10.sup.7.5) per strain is typical.
[0111] Influenza strains used with the invention may have a natural
HA as found in a wild-type virus, or a modified HA. For instance,
it is known to modify HA to remove determinants (e.g. hyper-basic
regions around the HA1/HA2 cleavage site) that cause a virus to be
highly pathogenic in avian species. The use of reverse genetics
facilitates such modifications.
[0112] As well as being suitable for immunizing against
inter-pandemic strains, the compositions of the invention are
particularly useful for immunizing against pandemic or
potentially-pandemic strains. The invention is suitable for
vaccinating humans as well as non-human animals
[0113] Other strains whose antigens can usefully be included in the
compositions are strains which are resistant to antiviral therapy
(e.g. resistant to oseltamivir [31] and/or zanamivir), including
resistant pandemic strains [32].
[0114] Compositions of the invention may include antigen(s) from
one or more (e.g. 1, 2, 3, 4 or more) influenza virus strains,
including influenza A virus and/or influenza B virus provided that
at least one influenza strain is a reassortant influenza strain of
the invention. Compositions wherein at least two, at least three or
all of the antigens are from reassortant influenza strains of the
invention are also envisioned. Where a vaccine includes more than
one strain of influenza, the different strains are typically grown
separately and are mixed after the viruses have been harvested and
antigens have been prepared. Thus a process of the invention may
include the step of mixing antigens from more than one influenza
strain. A trivalent vaccine is typical, including antigens from two
influenza A virus strains and one influenza B virus strain. A
tetravalent vaccine is also useful [33], including antigens from
two influenza A virus strains and two influenza B virus strains, or
three influenza A virus strains and one influenza B virus
strain.
Pharmaceutical Compositions
[0115] Vaccine compositions manufactured according to the invention
are pharmaceutically acceptable. They usually include components in
addition to the antigens e.g. they typically include one or more
pharmaceutical carrier(s) and/or excipient(s). As described below,
adjuvants may also be included. A thorough discussion of such
components is available in reference 34.
[0116] Vaccine compositions will generally be in aqueous form.
However, some vaccines may be in dry form, e.g. in the form of
injectable solids or dried or polymerized preparations on a
patch.
[0117] Vaccine compositions may include preservatives such as
thiomersal or 2-phenoxyethanol. It is preferred, however, that the
vaccine should be substantially free from (i.e. less than 5
.mu.g/ml) mercurial material e.g. thiomersal-free [Error! Bookmark
not defined, 35]. Vaccines containing no mercury are more
preferred. An .alpha.-tocopherol succinate can be included as an
alternative to mercurial compounds [Error! Bookmark not defined.].
Preservative-free vaccines are particularly preferred.
[0118] To control tonicity, it is preferred to include a
physiological salt, such as a sodium salt. Sodium chloride (NaCl)
is preferred, which may be present at between 1 and 20 mg/ml. Other
salts that may be present include potassium chloride, potassium
dihydrogen phosphate, disodium phosphate dehydrate, magnesium
chloride, calcium chloride, etc.
[0119] Vaccine compositions will generally have an osmolality of
between 200 mOsm/kg and 400 mOsm/kg, preferably between 240-360
mOsm/kg, and will more preferably fall within the range of 290-310
mOsm/kg. Osmolality has previously been reported not to have an
impact on pain caused by vaccination [36], but keeping osmolality
in this range is nevertheless preferred.
[0120] Vaccine compositions may include one or more buffers.
Typical buffers include: a phosphate buffer; a Tris buffer; a
borate buffer; a succinate buffer; a histidine buffer (particularly
with an aluminum hydroxide adjuvant); or a citrate buffer. Buffers
will typically be included in the 5-20 mM range.
[0121] The pH of a vaccine composition will generally be between
5.0 and 8.1, and more typically between 6.0 and 8.0 e.g. 6.5 and
7.5, or between 7.0 and 7.8. A process of the invention may
therefore include a step of adjusting the pH of the bulk vaccine
prior to packaging.
[0122] The vaccine composition is preferably sterile. The vaccine
composition is preferably non-pyrogenic e.g. containing <1 EU
(endotoxin unit, a standard measure) per dose, and preferably
<0.1 EU per dose. The vaccine composition is preferably
gluten-free.
[0123] Vaccine compositions of the invention may include detergent
e.g. a polyoxyethylene sorbitan ester surfactant (known as
`Tweens`), an octoxynol (such as octoxynol-9 (Triton X-100) or
t-octylphenoxypolyethoxyethanol), a cetyl trimethyl ammonium
bromide (`CTAB`), or sodium deoxycholate, particularly for a split
or surface antigen vaccine. The detergent may be present only at
trace amounts. Thus the vaccine may include less than 1 mg/ml of
each of octoxynol-10 and polysorbate 80. Other residual components
in trace amounts could be antibiotics (e.g. neomycin, kanamycin,
polymyxin B).
[0124] A vaccine composition may include material for a single
immunisation, or may include material for multiple immunisations
(i.e. a `multidose` kit). The inclusion of a preservative is
preferred in multidose arrangements. As an alternative (or in
addition) to including a preservative in multidose compositions,
the compositions may be contained in a container having an aseptic
adaptor for removal of material.
[0125] Influenza vaccines are typically administered in a dosage
volume of about 0.5 ml, although a half dose (i.e. about 0.25 ml)
may be administered to children.
[0126] Compositions and kits are preferably stored at between
2.degree. C. and 8.degree. C. They should not be frozen. They
should ideally be kept out of direct light.
Host Cell DNA
[0127] Where virus has been isolated and/or grown on a cell line,
it is standard practice to minimize the amount of residual cell
line DNA in the final vaccine, in order to minimize any potential
oncogenic activity of the DNA.
[0128] Thus a vaccine composition prepared according to the
invention preferably contains less than 10 ng (preferably less than
1 ng, and more preferably less than 100 pg) of residual host cell
DNA per dose, although trace amounts of host cell DNA may be
present.
[0129] It is preferred that the average length of any residual host
cell DNA is less than 500 bp e.g. less than 400 bp, less than 300
bp, less than 200 bp, less than 100 bp, etc.
[0130] Contaminating DNA can be removed during vaccine preparation
using standard purification procedures e.g. chromatography, etc.
Removal of residual host cell DNA can be enhanced by nuclease
treatment e.g. by using a DNase. A convenient method for reducing
host cell DNA contamination is disclosed in references 37 & 38,
involving a two-step treatment, first using a DNase (e.g.
Benzonase), which may be used during viral growth, and then a
cationic detergent (e.g. CTAB), which may be used during virion
disruption. Treatment with an alkylating agent, such as
.beta.-propiolactone, can also be used to remove host cell DNA, and
advantageously may also be used to inactivate virions [39].
Adjuvants
[0131] Compositions of the invention may advantageously include an
adjuvant, which can function to enhance the immune responses
(humoral and/or cellular) elicited in a subject who receives the
composition. Preferred adjuvants comprise oil-in-water emulsions.
Various such adjuvants are known, and they typically include at
least one oil and at least one surfactant, with the oil(s) and
surfactant(s) being biodegradable (metabolisable) and
biocompatible. The oil droplets in the emulsion are generally less
than 5 .mu.m in diameter, and ideally have a sub-micron diameter,
with these small sizes being achieved with a microfluidiser to
provide stable emulsions. Droplets with a size less than 220 nm are
preferred as they can be subjected to filter sterilization.
[0132] The emulsion can comprise oils such as those from an animal
(such as fish) or vegetable source. Sources for vegetable oils
include nuts, seeds and grains. Peanut oil, soybean oil, coconut
oil, and olive oil, the most commonly available, exemplify the nut
oils. Jojoba oil can be used e.g. obtained from the jojoba bean.
Seed oils include safflower oil, cottonseed oil, sunflower seed
oil, sesame seed oil and the like. In the grain group, corn oil is
the most readily available, but the oil of other cereal grains such
as wheat, oats, rye, rice, teff, triticale and the like may also be
used. 6-10 carbon fatty acid esters of glycerol and
1,2-propanediol, while not occurring naturally in seed oils, may be
prepared by hydrolysis, separation and esterification of the
appropriate materials starting from the nut and seed oils. Fats and
oils from mammalian milk are metabolizable and may therefore be
used in the practice of this invention. The procedures for
separation, purification, saponification and other means necessary
for obtaining pure oils from animal sources are well known in the
art. Most fish contain metabolizable oils which may be readily
recovered. For example, cod liver oil, shark liver oils, and whale
oil such as spermaceti exemplify several of the fish oils which may
be used herein. A number of branched chain oils are synthesized
biochemically in 5-carbon isoprene units and are generally referred
to as terpenoids. Shark liver oil contains a branched, unsaturated
terpenoids known as squalene,
2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, which
is particularly preferred herein. Squalane, the saturated analog to
squalene, is also a preferred oil. Fish oils, including squalene
and squalane, are readily available from commercial sources or may
be obtained by methods known in the art. Another preferred oil is
.alpha.-tocopherol (see below).
[0133] Mixtures of oils can be used.
[0134] Surfactants can be classified by their `HLB`
(hydrophile/lipophile balance). Preferred surfactants of the
invention have a HLB of at least 10, preferably at least 15, and
more preferably at least 16. The invention can be used with
surfactants including, but not limited to: the polyoxyethylene
sorbitan esters surfactants (commonly referred to as the Tweens),
especially polysorbate 20 and polysorbate 80; copolymers of
ethylene oxide (E0), propylene oxide (PO), and/or butylene oxide
(BO), sold under the DOWFAX.TM. tradename, such as linear EO/PO
block copolymers; octoxynols, which can vary in the number of
repeating ethoxy(oxy-1,2-ethanediyl) groups, with octoxynol-9
(Triton X-100, or t-octylphenoxypolyethoxyethanol) being of
particular interest; (octylphenoxy)polyethoxyethanol (IGEPAL
CA-630/NP-40); phospholipids such as phosphatidylcholine
(lecithin); nonylphenol ethoxylates, such as the Tergitol.TM. NP
series; polyoxyethylene fatty ethers derived from lauryl, cetyl,
stearyl and oleyl alcohols (known as Brij surfactants), such as
triethyleneglycol monolauryl ether (Brij 30); and sorbitan esters
(commonly known as the SPANs), such as sorbitan trioleate (Span 85)
and sorbitan monolaurate. Non-ionic surfactants are preferred.
Preferred surfactants for including in the emulsion are Tween 80
(polyoxyethylene sorbitan monooleate), Span 85 (sorbitan
trioleate), lecithin and Triton X-100.
[0135] Mixtures of surfactants can be used e.g. Tween 80/Span 85
mixtures. A combination of a polyoxyethylene sorbitan ester such as
polyoxyethylene sorbitan monooleate (Tween 80) and an octoxynol
such as t-octylphenoxypolyethoxyethanol (Triton X-100) is also
suitable. Another useful combination comprises laureth 9 plus a
polyoxyethylene sorbitan ester and/or an octoxynol.
[0136] Preferred amounts of surfactants (% by weight) are:
polyoxyethylene sorbitan esters (such as Tween 80) 0.01 to 1%, in
particular about 0.1%; octyl- or nonylphenoxy polyoxyethanols (such
as Triton X-100, or other detergents in the Triton series) 0.001 to
0.1%, in particular 0.005 to 0.02%; polyoxyethylene ethers (such as
laureth 9) 0.1 to 20%, preferably 0.1 to 10% and in particular 0.1
to 1% or about 0.5%.
[0137] Where the vaccine contains a split virus, it is preferred
that it contains free surfactant in the aqueous phase. This is
advantageous as the free surfactant can exert a `splitting effect`
on the antigen, thereby disrupting any unsplit virions and/or
virion aggregates that might otherwise be present. This can improve
the safety of split virus vaccines [40].
[0138] Preferred emulsions have an average droplets size of <1
.mu.m e.g. .ltoreq.750 nm, .ltoreq.500 nm, .ltoreq.400 nm,
.ltoreq.300 nm, .ltoreq.250 nm, .ltoreq.220 nm, .ltoreq.200 nm, or
smaller. These droplet sizes can conveniently be achieved by
techniques such as microfluidisation.
[0139] Specific oil-in-water emulsion adjuvants useful with the
invention include, but are not limited to: [0140] A submicron
emulsion of squalene, Tween 80, and Span 85. The composition of the
emulsion by volume can be about 5% squalene, about 0.5% polysorbate
80 and about 0.5% Span 85. In weight terms, these ratios become
4.3% squalene, 0.5% polysorbate 80 and 0.48% Span 85. This adjuvant
is known as `MF59` [41-42], as described in more detail in Chapter
10 of ref. 43 and chapter 12 of ref. 44. The MF59 emulsion
advantageously includes citrate ions e.g. 10 mM sodium citrate
buffer. [0141] An emulsion comprising squalene, a tocopherol, and
polysorbate 80. The emulsion may include phosphate buffered saline.
These emulsions may have by volume from 2 to 10% squalene, from 2
to 10% tocopherol and from 0.3 to 3% polysorbate 80, and the weight
ratio of squalene:tocopherol is preferably <1 (e.g. 0.90) as
this can provide a more stable emulsion. Squalene and polysorbate
80 may be present volume ratio of about 5:2 or at a weight ratio of
about 11:5. Thus the three components (squalene, tocopherol,
polysorbate 80) may be present at a weight ratio of 1068:1186:485
or around 55:61:25. One such emulsion (`AS03`) can be made by
dissolving Tween 80 in PBS to give a 2% solution, then mixing 90 ml
of this solution with a mixture of (5 g of DL a tocopherol and 5 ml
squalene), then microfluidising the mixture. The resulting emulsion
may have submicron oil droplets e.g. with an average diameter of
between 100 and 250 nm, preferably about 180 nm. The emulsion may
also include a 3-de-O-acylated monophosphoryl lipid A (3d MPL).
Another useful emulsion of this type may comprise, per human dose,
0.5-10 mg squalene, 0.5-11 mg tocopherol, and 0.1-4 mg polysorbate
80 [45] e.g. in the ratios discussed above. [0142] An emulsion of
squalene, a tocopherol, and a Triton detergent (e.g. Triton X-100).
The emulsion may also include a 3d-MPL (see below). The emulsion
may contain a phosphate buffer. [0143] An emulsion comprising a
polysorbate (e.g. polysorbate 80), a Triton detergent (e.g. Triton
X-100) and a tocopherol (e.g. an .alpha.-tocopherol succinate). The
emulsion may include these three components at a mass ratio of
about 75:11:10 (e.g. 750 .mu.g/ml polysorbate 80, 110 .mu.g/ml
Triton X-100 and 100 .mu.g/ml .alpha.-tocopherol succinate), and
these concentrations should include any contribution of these
components from antigens. The emulsion may also include squalene.
The emulsion may also include a 3d-MPL (see below). The aqueous
phase may contain a phosphate buffer. [0144] An emulsion of
squalane, polysorbate 80 and poloxamer 401 ("Pluronic.TM. L121").
The emulsion can be formulated in phosphate buffered saline, pH
7.4. This emulsion is a useful delivery vehicle for muramyl
dipeptides, and has been used with threonyl-MDP in the "SAF-1"
adjuvant [46] (0.05-1% Thr-MDP, 5% squalane, 2.5% Pluronic L121 and
0.2% polysorbate 80). It can also be used without the Thr-MDP, as
in the "AF" adjuvant [47] (5% squalane, 1.25% Pluronic L121 and
0.2% polysorbate 80). Microfluidisation is preferred. [0145] An
emulsion comprising squalene, an aqueous solvent, a polyoxyethylene
alkyl ether hydrophilic nonionic surfactant (e.g. polyoxyethylene
(12) cetostearyl ether) and a hydrophobic nonionic surfactant (e.g.
a sorbitan ester or mannide ester, such as sorbitan monoleate or
`Span 80`). The emulsion is preferably thermoreversible and/or has
at least 90% of the oil droplets (by volume) with a size less than
200 nm [48]. The emulsion may also include one or more of: alditol;
a cryoprotective agent (e.g. a sugar, such as dodecylmaltoside
and/or sucrose); and/or an alkylpolyglycoside. The emulsion may
include a TLR4 agonist [49]. Such emulsions may be lyophilized.
[0146] An emulsion of squalene, poloxamer 105 and Abil-Care [50].
The final concentration (weight) of these components in adjuvanted
vaccines are 5% squalene, 4% poloxamer 105 (pluronic polyol) and 2%
Abil-Care 85 (Bis-PEG/PPG-16/16 PEG/PPG-16/16 dimethicone;
caprylic/capric triglyceride). [0147] An emulsion having from
0.5-50% of an oil, 0.1-10% of a phospholipid, and 0.05-5% of a
non-ionic surfactant. As described in reference 51, preferred
phospholipid components are phosphatidylcholine,
phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol,
phosphatidylglycerol, phosphatidic acid, sphingomyelin and
cardiolipin. Submicron droplet sizes are advantageous. [0148] A
submicron oil-in-water emulsion of a non-metabolisable oil (such as
light mineral oil) and at least one surfactant (such as lecithin,
Tween 80 or Span 80). Additives may be included, such as QuilA
saponin, cholesterol, a saponin-lipophile conjugate (such as
GPI-0100, described in reference 52, produced by addition of
aliphatic amine to desacylsaponin via the carboxyl group of
glucuronic acid), dimethyldioctadecylammonium bromide and/or
N,N-dioctadecyl-N,N-bis(2-hydroxyethyl)propanediamine. [0149] An
emulsion in which a saponin (e.g. QuilA or QS21) and a sterol (e.g.
a cholesterol) are associated as helical micelles [53]. [0150] An
emulsion comprising a mineral oil, a non-ionic lipophilic
ethoxylated fatty alcohol, and a non-ionic hydrophilic surfactant
(e.g. an ethoxylated fatty alcohol and/or
polyoxyethylene-polyoxypropylene block copolymer) [54]. [0151] An
emulsion comprising a mineral oil, a non-ionic hydrophilic
ethoxylated fatty alcohol, and a non-ionic lipophilic surfactant
(e.g. an ethoxylated fatty alcohol and/or
polyoxyethylene-polyoxypropylene block copolymer) [54].
[0152] In some embodiments an emulsion may be mixed with antigen
extemporaneously, at the time of delivery, and thus the adjuvant
and antigen may be kept separately in a packaged or distributed
vaccine, ready for final formulation at the time of use. In other
embodiments an emulsion is mixed with antigen during manufacture,
and thus the composition is packaged in a liquid adjuvanted form.
The antigen will generally be in an aqueous form, such that the
vaccine is finally prepared by mixing two liquids. The volume ratio
of the two liquids for mixing can vary (e.g. between 5:1 and 1:5)
but is generally about 1:1. Where concentrations of components are
given in the above descriptions of specific emulsions, these
concentrations are typically for an undiluted composition, and the
concentration after mixing with an antigen solution will thus
decrease.
Packaging of Vaccine Compositions
[0153] Suitable containers for compositions of the invention (or
kit components) include vials, syringes (e.g. disposable syringes),
nasal sprays, etc. These containers should be sterile.
[0154] Where a composition/component is located in a vial, the vial
is preferably made of a glass or plastic material. The vial is
preferably sterilized before the composition is added to it. To
avoid problems with latex-sensitive patients, vials are preferably
sealed with a latex-free stopper, and the absence of latex in all
packaging material is preferred. The vial may include a single dose
of vaccine, or it may include more than one dose (a `multidose`
vial) e.g. 10 doses. Preferred vials are made of colourless
glass.
[0155] A vial can have a cap (e.g. a Luer lock) adapted such that a
pre-filled syringe can be inserted into the cap, the contents of
the syringe can be expelled into the vial (e.g. to reconstitute
lyophilised material therein), and the contents of the vial can be
removed back into the syringe. After removal of the syringe from
the vial, a needle can then be attached and the composition can be
administered to a patient. The cap is preferably located inside a
seal or cover, such that the seal or cover has to be removed before
the cap can be accessed. A vial may have a cap that permits aseptic
removal of its contents, particularly for multidose vials.
[0156] Where a component is packaged into a syringe, the syringe
may have a needle attached to it. If a needle is not attached, a
separate needle may be supplied with the syringe for assembly and
use. Such a needle may be sheathed. Safety needles are preferred.
1-inch 23-gauge, 1-inch 25-gauge and 5/8-inch 25-gauge needles are
typical. Syringes may be provided with peel-off labels on which the
lot number, influenza season and expiration date of the contents
may be printed, to facilitate record keeping. The plunger in the
syringe preferably has a stopper to prevent the plunger from being
accidentally removed during aspiration. The syringes may have a
latex rubber cap and/or plunger. Disposable syringes contain a
single dose of vaccine. The syringe will generally have a tip cap
to seal the tip prior to attachment of a needle, and the tip cap is
preferably made of a butyl rubber. If the syringe and needle are
packaged separately then the needle is preferably fitted with a
butyl rubber shield. Preferred syringes are those marketed under
the trade name "Tip-Lok".TM..
[0157] Containers may be marked to show a half-dose volume e.g. to
facilitate delivery to children. For instance, a syringe containing
a 0.5 ml dose may have a mark showing a 0.25 ml volume.
[0158] Where a glass container (e.g. a syringe or a vial) is used,
then it is preferred to use a container made from a borosilicate
glass rather than from a soda lime glass.
[0159] A kit or composition may be packaged (e.g. in the same box)
with a leaflet including details of the vaccine e.g. instructions
for administration, details of the antigens within the vaccine,
etc. The instructions may also contain warnings e.g. to keep a
solution of adrenaline readily available in case of anaphylactic
reaction following vaccination, etc.
Methods of Treatment, and Administration of the Vaccine
[0160] The invention provides a vaccine manufactured according to
the invention. These vaccine compositions are suitable for
administration to human or non-human animal subjects, such as pigs
or birds, and the invention provides a method of raising an immune
response in a subject, comprising the step of administering a
composition of the invention to the subject. The invention also
provides a composition of the invention for use as a medicament,
and provides the use of a composition of the invention for the
manufacture of a medicament for raising an immune response in a
subject.
[0161] The immune response raised by these methods and uses will
generally include an antibody response, preferably a protective
antibody response. Methods for assessing antibody responses,
neutralising capability and protection after influenza virus
vaccination are well known in the art. Human studies have shown
that antibody titers against hemagglutinin of human influenza virus
are correlated with protection (a serum sample
hemagglutination-inhibition titer of about 30-40 gives around 50%
protection from infection by a homologous virus) [55]. Antibody
responses are typically measured by hemagglutination inhibition, by
microneutralisation, by single radial immunodiffusion (SRID),
and/or by single radial hemolysis (SRH). These assay techniques are
well known in the art.
[0162] Compositions of the invention can be administered in various
ways. The most preferred immunisation route is by intramuscular
injection (e.g. into the arm or leg), but other available routes
include subcutaneous injection, intranasal [56-57], oral [58],
intradermal [59,60], transcutaneous, transdermal [61], etc.
[0163] Vaccines prepared according to the invention may be used to
treat both children and adults. Influenza vaccines are currently
recommended for use in pediatric and adult immunisation, from the
age of 6 months. Thus a human subject may be less than 1 year old,
1-5 years old, 5-15 years old, 15-55 years old, or at least 55
years old. Preferred subjects for receiving the vaccines are the
elderly (e.g. >50 years old, .gtoreq.60 years old, and
preferably .gtoreq.65 years), the young (e.g. .ltoreq.5 years old),
hospitalised subjects, healthcare workers, armed service and
military personnel, pregnant women, the chronically ill,
immunodeficient subjects, subjects who have taken an antiviral
compound (e.g. an oseltamivir or zanamivir compound; see below) in
the 7 days prior to receiving the vaccine, people with egg
allergies and people travelling abroad. The vaccines are not
suitable solely for these groups, however, and may be used more
generally in a population. For pandemic strains, administration to
all age groups is preferred.
[0164] Preferred compositions of the invention satisfy 1, 2 or 3 of
the CPMP criteria for efficacy. In adults (18-60 years), these
criteria are: (1) .gtoreq.70% seroprotection; (2) .gtoreq.40%
seroconversion; and/or (3) a GMT increase of .gtoreq.2.5-fold. In
elderly (>60 years), these criteria are: (1) .gtoreq.60%
seroprotection; (2) .gtoreq.30% seroconversion; and/or (3) a GMT
increase of .gtoreq.2-fold. These criteria are based on open label
studies with at least 50 patients.
[0165] Treatment can be by a single dose schedule or a multiple
dose schedule. Multiple doses may be used in a primary immunisation
schedule and/or in a booster immunisation schedule. In a multiple
dose schedule the various doses may be given by the same or
different routes e.g. a parenteral prime and mucosal boost, a
mucosal prime and parenteral boost, etc. Administration of more
than one dose (typically two doses) is particularly useful in
immunologically naive patients e.g. for people who have never
received an influenza vaccine before, or for vaccinating against a
new HA subtype (as in a pandemic outbreak). Multiple doses will
typically be administered at least 1 week apart (e.g. about 2
weeks, about 3 weeks, about 4 weeks, about 6 weeks, about 8 weeks,
about 10 weeks, about 12 weeks, about 16 weeks, etc.).
[0166] Vaccines produced by the invention may be administered to
patients at substantially the same time as (e.g. during the same
medical consultation or visit to a healthcare professional or
vaccination centre) other vaccines e.g. at substantially the same
time as a measles vaccine, a mumps vaccine, a rubella vaccine, a
MMR vaccine, a varicella vaccine, a MMRV vaccine, a diphtheria
vaccine, a tetanus vaccine, a pertussis vaccine, a DTP vaccine, a
conjugated H. influenzae type b vaccine, an inactivated poliovirus
vaccine, a hepatitis B virus vaccine, a meningococcal conjugate
vaccine (such as a tetravalent A-C-W135-Y vaccine), a respiratory
syncytial virus vaccine, a pneumococcal conjugate vaccine, etc.
Administration at substantially the same time as a pneumococcal
vaccine and/or a meningococcal vaccine is particularly useful in
elderly patients.
[0167] Similarly, vaccines of the invention may be administered to
patients at substantially the same time as (e.g. during the same
medical consultation or visit to a healthcare professional) an
antiviral compound, and in particular an antiviral compound active
against influenza virus (e.g. oseltamivir and/or zanamivir). These
antivirals include neuraminidase inhibitors, such as a
(3R,4R,5S)-4-acetylamino-5-amino-3(1-ethylpropoxy)-1-cyclohexene-1-carbox-
ylic acid or
5-(acetylamino)-4-[(aminoiminomethyl)-amino]-2,6-anhydro-3,4,5-trideoxy-D-
-glycero-D-galactonon-2-enonic acid, including esters thereof (e.g.
the ethyl esters) and salts thereof (e.g. the phosphate salts). A
preferred antiviral is
(3R,4R,5S)-4-acetylamino-5-amino-3(1-ethylpropoxy)-1-cyclohexene-1-carbox-
ylic acid, ethyl ester, phosphate (1:1), also known as oseltamivir
phosphate (TAMIFLU.TM.).
General
[0168] The term "comprising" encompasses "including" as well as
"consisting" e.g. a composition "comprising" X may consist
exclusively of X or may include something additional e.g. X+Y.
[0169] The word "substantially" does not exclude "completely" e.g.
a composition which is "substantially free" from Y may be
completely free from Y. Where necessary, the word "substantially"
may be omitted from the definition of the invention.
[0170] The term "about" in relation to a numerical value x is
optional and means, for example, x.+-.10%.
[0171] Unless specifically stated, a process comprising a step of
mixing two or more components does not require any specific order
of mixing. Thus components can be mixed in any order. Where there
are three components then two components can be combined with each
other, and then the combination may be combined with the third
component, etc.
[0172] The various steps of the methods may be carried out at the
same or different times, in the same or different geographical
locations, e.g. countries, and by the same or different people or
entities.
[0173] Where animal (and particularly bovine) materials are used in
the culture of cells, they should be obtained from sources that are
free from transmissible spongiform encephalopathies (TSEs), and in
particular free from bovine spongiform encephalopathy (BSE).
Overall, it is preferred to culture cells in the total absence of
animal-derived materials.
[0174] Where a compound is administered to the body as part of a
composition then that compound may alternatively be replaced by a
suitable prodrug.
[0175] References to a percentage sequence identity between two
amino acid sequences means that, when aligned, that percentage of
amino acids are the same in comparing the two sequences. This
alignment and the percent homology or sequence identity can be
determined using software programs known in the art, for example
those described in section 7.7.18 of reference 62. A preferred
alignment is determined by the Smith-Waterman homology search
algorithm using an affine gap search with a gap open penalty of 12
and a gap extension penalty of 2, BLOSUM matrix of 62. The
Smith-Waterman homology search algorithm is taught in reference
63.
[0176] References to a percentage sequence identity between two
nucleic acid sequences mean that, when aligned, that percentage of
bases are the same in comparing the two sequences. This alignment
and the percent homology or sequence identity can be determined
using software programs known in the art, for example those
described in section 7.7.18 of reference 62. A preferred alignment
program is GCG Gap (Genetics Computer Group, Wisconsin, Suite
Version 10.1), preferably using default parameters, which are as
follows: open gap=3; extend gap=1.
BRIEF DESCRIPTION OF THE DRAWINGS
[0177] FIG. 1 illustrates virus titers (by Focus-Formation assay
(FFA); (A) and HA titers (by Red Blood Cell Hemagglutination assay;
(B) at different times post-infection of wt PR8 and PR8-X viruses
grown in MDCK cells. The solid line in (A) and hatched columns in
(B) represent results with wild-type PR8. The dotted line in (A)
and empty columns in (B) represent results with wild-type PR8-X.
The x-axis shows the hours post infection and the y-axis in (A) and
(B) shows the virus titer (IU/ml) and HA titre, respectively.
[0178] FIG. 2 illustrates virus titers (by FFA; (A) and HA titers
(by Red Blood Cell Hemagglutination assay; (B) at different times
post-infection of reverse genetics derived PR8 and PR8-X viruses
grown in MDCK cells. The solid line in (A) and hatched columns in
(B) represent results with PR8. The dotted line in (A) and empty
columns in (B) represent results with RG-derived PR8-X. The x-axis
shows the hours post infection and the y-axis in (A) and (B) shows
the virus titer (IU/ml) and HA titre, respectively.
[0179] FIG. 3 compares virus titers (by FFA; (A) and HA titers (by
Red Blood Cell Hemagglutination assay; (B)--at different times
post-infection in MDCK cells of reverse genetics-derived 6:2
reassortant viruses made with either PR8 or PR8-X backbone segments
which contain the HA and NA segments from PR8-X. The solid line in
(A) and hatched columns in (B) represent results with the PR8
backbone. The dotted line in (A) and empty columns in (B) represent
results with the PR8-X backbone. The x-axis shows the hours post
infection and the y-axis in (A) and (B) shows the virus titer
(IU/ml) and HA titre, respectively.
[0180] FIG. 4 compares virus titers by FFA (A) and HA titers (by
Red Blood Cell Hemagglutination assay; (B) at different times
post-infection in MDCK cells of reverse genetics-derived 6:2
reassortant viruses made with either wt PR8 or PR8-X backbone
segments which contain the HA and NA segments from a pandemic H1
strain (strain 1). The solid line in (A) and hatched columns in (B)
represent results with the wt PR8 backbone. The dotted line in (A)
and empty columns in (B) represent results with the PR8-X backbone.
The x-axis shows the hours post infection and the y-axis in (A) and
(B) shows the virus titer (IU/ml) and HA titre, respectively.
[0181] FIG. 5 compares virus titers by a focus-formation assay
(FFA) (A) and HA titers (B) at different times post-infection in
MDCK cells of reverse genetics-derived 6:2 reassortant viruses made
with either PR8 or PR8-X backbone segments which contain the HA and
NA segments from 105p30. The solid line in (A) and hatched columns
in (B) represent results with the wt PR8 backbone. The dotted line
in (A) and empty columns in (B) represent results with the PR8-X
backbone. The x-axis shows the hours post infection and the y-axis
shows the virus titer (IU/ml).
[0182] FIG. 6 illustrates virus titers by a focus-formation assay
(FFA) at different times post-infection of wild-type PR8-X and
105p30 viruses (A) or reverse genetics-derived PR8-X and 105p30
viruses (B) grown in MDCK cells. In (A) and (B), the solid lines
represent results with 105p30. The dotted lines represent results
with PR8-X. The x-axis shows the hours post infection and the
y-axis in (A) and (B) shows the virus titer (IU/ml) and HA titre,
respectively.
[0183] FIG. 7 shows the growth characteristics of reassortant
viruses containing the backbone segments of the wt PR8 strain (line
with triangles) or 105p30 strain (line with squares) and the HA and
NA segments of a pandemic H1 influenza strain (strain 2). The
x-axis in (A) and (B) indicates the hours post infection. The
y-axis in (A) shows the titre Log 10 in FFU per mL. The y-axis in
(B) shows the titre log 10 in virus particles per mL.
[0184] FIG. 8 compares virus titers by a focus-formation assay
(FFA) at different times post-infection in MDCK cells of reverse
genetics-derived 6:2 reassortant viruses made with either 105p30 or
PR8-X backbone segments which contain the HA and NA segments from
(A) a H1 strain (strain 1) or (B) a pandemic H1 strain (strain 2).
The solid lines represent results with the 105p30 backbone. The
dotted lines represent results with the PR8-X backbone. The x-axis
shows the hours post infection and the y-axis shows the virus titer
(IU/ml).
[0185] FIG. 9 compares virus titers by a focus-formation assay
(FFA) at different times post-infection in MDCK cells of reverse
genetics-derived 6:2 reassortant viruses made with either the #17,
#19, or PR8-X backbone in combination with the HA and NA segments
from (A) a pandemic H1 strain (strain 3) or (B) a H3 (strain 1). In
(A) and (B), the dotted lines with the circle markers represent
results with the #17 backbone. The solid lines with diamond markers
represent results with the #19 backbone. The dotted lines with
square markers represent results with the PR8-X backbone. The
x-axis shows the hours post infection and the y-axis shows the
virus titer (IU/ml).
[0186] FIG. 10 compares virus titers by a focus-formation assay
(FFA) at different times post-infection in MDCK cells of a panel of
different reverse genetics-derived 6:2 reassortant viruses made
with either the chimeric #19 or PR8-X backbone plus the HA and NA
segments from the following strains: (A) a pandemic H1 strain
(strain 2), (B) a pandemic H1 strain (strain 4)-In (A) and (B), the
solid lines with the triangle markers represent results with the
#19 backbone. The dotted lines with square markers represent
results with the PR8-X backbone. The x-axis shows the hours post
infection and the y-axis shows the virus titer (IU/ml).
[0187] FIG. 11 compares virus titers by a focus-formation assay
(FFA) at different times post-infection in MDCK cells of a panel of
different reverse genetics-derived 6:2 reassortant viruses made
with either the chimeric #19 or PR8-X backbone plus the HA and NA
segments from the following strains: (C) a H1 strain (strain 2),
(D) a H1 strain (strain 3). In (C) and (D), the solid lines with
the triangle markers represent results with the #19 backbone. The
dotted lines with square markers represent results with the PR8-X
backbone. The x-axis shows the hours post infection and the y-axis
shows the virus titer (IU/ml).
[0188] FIG. 12 compares virus titers by a focus-formation assay
(FFA) at different times post-infection in MDCK cells of a panel of
different reverse genetics-derived 6:2 reassortant viruses made
with either the chimeric #19 or PR8-X backbone plus the HA and NA
segments from the following strain: a H3 strain (strain 2). In FIG.
12, the solid lines with the triangle markers represent results
with the #19 backbone. The dotted lines with square markers
represent results with the PR8-X backbone. The x-axis shows the
hours post infection and the y-axis shows the virus titer
(IU/ml).
[0189] FIG. 13 compares HA yields (by lectin-capture ELISA) at 60
hr post-infection in MDCK cells of different 6:2 reassortant
viruses made with either the chimeric #19 (empty columns) or PR8-X
backbone (solid columns) plus the HA and NA segments from the
following strains: (A) a pandemic H1 strain (strain 2), (B) a
pandemic H1 strain (strain 4). Corresponding 6:2 reassortant
viruses made by classical reassortment ("classical") with the wt
PR8 backbone were included as controls (hatched columns). The
y-axis shows the HA content in .mu.g per mL.
[0190] FIG. 14 compares HA yields (by lectin-capture ELISA) at 60
hr post-infection in MDCK cells of different 6:2 reassortant
viruses made with either the chimeric #19 (empty columns) or PR8-X
backbone (solid columns) plus the HA and NA segments from the
following strains: (C) a H3 strain (strain 1), or (D) a H3 strain
(strain 2). Corresponding 6:2 reassortant viruses made by classical
reassortment ("classical") with the wt PR8 backbone were included
as controls (hatched columns) The y-axis shows the HA content in
.mu.g per mL.
[0191] FIG. 15 shows the growth curves of reassortant influenza
viruses. (A) shows growth curves of reassortant influenza viruses
comprising backbones 17, 18, 19 and 20 (as shown in table 1; line
with diamonds, squares, triangles and crosses, respectively), a
control comprising the same HA and NA segments from a H3 influenza
strain (strain 1) but all backbone segments from PR8-X (line with
circles) and the equivalent wildtype strain (line with plus sign).
The x axis indicates the hours post infection (hpi) and the y-axis
shows IU/mL. (B) shows the growth curve of reassortant influenza
viruses comprising backbones 17 and 19 (line with diamonds and
triangles, respectively) and the HA segments from a H3 influenza
strain (strain 3), a control comprising the same HA and NA segments
but all backbone segments from PR8-X (line with plus sign) and the
equivalent wildtype strain (line with circles).
[0192] FIG. 16 shows the results of a FFA (A) and HA-ELISA (B)
assay using reassortant influenza viruses comprising backbone 19
(open box), PR8-X backbone (hatched box) and the wildtype influenza
virus (dotted box). (A) and (B) show the results with a H1
influenza strain (strain 2). The y axis in (A) indicates the virus
titre in IU/mL and the y axis in (B) indicates HA in .mu.g/mL.
[0193] FIG. 17 shows the results of a FFA (C) and HA-ELISA (D)
assay using reassortant influenza viruses comprising backbone 19
(open box), PR8-X backbone (hatched box) and the wildtype influenza
virus (dotted box). (C) and (D) show the results with a H3
influenza virus strain. The y axis in (C) indicates the virus titre
in IU/mL and the y axis in (D) indicates HA in .mu.g/mL.
[0194] FIG. 18 is an alignment of the M1 viral segment of A/New
Caledonia/20/99 (SEQ ID NO: 33) and 105p30 (SEQ ID NO: 45).
MODES FOR CARRYING OUT THE INVENTION
Development of New Donor Strains
[0195] In order to provide high-growth donor strains, the donor
strain A/Puerto Rico/8/34 is passaged in MDCK 33016 cells five
times. Using this method, the inventors were able to obtain the
strain PR8-X which shows improved growth characteristics compared
with the original strain.
[0196] The 105p30 influenza donor strain was provided by isolating
an A/New Caledonia/20/1999 influenza virus from a clinical isolate
in MDCK 33016 cells and passaging the virus 30 times. The resulting
strain has a M segment with lysine in the position corresponding to
amino acid 95 of SEQ ID NO: 33 when aligned to SEQ ID NO: 33.
Growth Characteristics of Wt PR8 and PR8-X Viruses
[0197] In order to compare the growth characteristics of PR8-X and
wt PR8 donor strains, the viral titre of these virus strains is
measured in MDCK cells by focus-forming assays and hemagglutination
assays.
Focus-Forming Assays (FFA)
[0198] For the FFA, uninfected MDCK cells are plated at a density
of 1.8.times.10.sup.4 cells/well in 96 well plates in 100 .mu.l of
DMEM with 10% FCS. The next day, medium is aspirated and cells are
infected with viruses in a volume of 50 .mu.l (viruses diluted in
DMEM+1% FCS). The cells are incubated at 37.degree. C. until the
next day.
[0199] At several time points after infection, the medium is
aspirated and the cells washed once with PBS. 50 .mu.l of ice-cold
50%/50% acetone-methanol is added to each well followed by
incubation at -20.degree. C. for 30 minutes. The acetone mix is
aspirated and the cells washed once with PBST (PBS+0.1% Tween). 50
.mu.l of 2% BSA in PBS is added to each well followed by incubation
at room temperature (RT) for 30 minutes. 50 .mu.l of a 1:6000
dilution of anti-NP is added in blocking buffer followed by
incubation at RT for 1 hours. The antibody solution is aspirated
and the cells washed three times with PBST. Secondary antibody
(goat anti mouse) is added at a dilution 1:2000 in 50 .mu.l
blocking buffer and the plate is incubated at RT for 1 hours. The
antibody solution is aspirated and the cells washed three times
with PBST. 50 .mu.l of KPL True Blue is added to each well and
incubated for 10 minutes. The reaction is stopped by aspirating the
True-Blue and washing once with dH.sub.2O. The water is aspirated
and the cells are left to dry.
[0200] The results (FIG. 1) show that the PR8-X strain can grow to
higher titres in the same time frame compared to the wt PR8 strain
from which it is derived.
Growth Characteristics of Reassortant Viruses Containing PR8-X or
Wt PR8 Backbones
[0201] In order to test the suitability of the PR8-X strain as a
donor strain for virus reassortment, reassortant viruses are
produced by reverse genetics which contain the HA and NA proteins
from a pandemic H1 strain and the other viral segments from either
PR8-X or PR8. The viral titres of these reassortant viruses are
determined by FFA and HA assays as described above. The results are
shown in FIG. 4.
[0202] The results indicate that reassortant viruses which contain
viral segments from PR8-X grow faster in MDCK cells compared to
reassortant viruses containing viral segments from the PR8/34
strain.
Growth Characteristics of 105p30 Strain Compared with PR8-X
[0203] MDCK cells are infected with 105p30 and PR8-X at a moi of
10.sup.-3 and samples are taken at several time points after
infection. The titre is determined by a FFA assay. The results show
that 105p30 grows even faster in MDCK cells compared to PR8-X (FIG.
6).
Growth Characteristics of Reassortant Viruses Containing 105p30 or
wt PR8 Backbones
[0204] In order to test the suitability of the 105p30 strain as a
donor strain for virus reassortment, reverse genetics is used to
produce reassortant viruses that contain the HA and NA segments
from a pandemic H1 influenza strain and the backbone segments
either from the 105p30 or the wt PR8 strain. MDCK cells are
infected with the reassortant viruses at a moi of 10.sup.-3 and
samples are taken 1 hour, 12 hours, 36 hours and 60 hours after
infection. The titres are determined either by focus-forming assays
or by determining the virus particles by real-time detection PCR.
The reassortant viruses that contain the backbone segments from the
105p30 strain grow faster than the viruses that are reassorted with
the backbone segments of the wt PR8 strain. This shows that the
105p30 strain is a good donor strain for producing fast-growing
reassortant viruses (FIG. 7).
Rescue of Influenza Viruses Using Backbone Segments from Two Donor
Strains
[0205] The rescue efficiency of reassortant influenza viruses
containing the HA and NA segments from a H3 influenza virus and
backbone segments from the 105p30 and the PR8-X donor strains is
tested in MDCK cells. The reassortant influenza viruses contain
backbone segments of the 105p30 and the PR8-X donor strains, as
indicated in the following table:
TABLE-US-00001 TABLE 1 Backbone # FB1 PB2 PA NP M NS 1 PR8-X PR8-X
PR8-X 105p30 105p30 105p30 2 PR8-X PR8-X 105p30 PR8-X 105p30 105p30
3 PR8-X PR8-X 105p30 105p30 PR8-X 105p30 4 PR8-X PR8-X 105p30
105p30 105p30 PR8-X 5 PR8-X 105p30 PR8-X PR8-X 105p30 105p30 6
PR8-X 105p30 PR8-X 105p30 PR8-X 105p30 7 PR8-X 105p30 PR8-X 105p30
105p30 PR8-X 8 PR8-X 105p30 105p30 PR8-X PR8-X 105p30 9 PR8-X
105p30 105p30 PR8-X 105p30 PR8-X 10 PR8-X 105p30 105p30 105p30
PR8-X PR8-X 11 105p30 PR8-X PR8-X PR8-X 105p30 105p30 12 105p30
PR8-X PR8-X 105p30 PR8-X 105p30 13 105p30 PR8-X PR8-X 105p30 105p30
PR8-X 14 105p30 PR8-X 105p30 PR8-X 105p30 PR8-X 15 105p30 PR8-X
105p30 PR8-X PR8-X 105p30 16 105p30 PR8-X 105p30 105p30 PR8-X PR8-X
17 105p30 105p30 PR8-X PR8-X PR8-X 105p30 18 105p30 105p30 PR8-X
PR8-X 105p30 PR8-X 19 105p30 105p30 PR8-X 105p30 PR8-X PR8-X 20
105p30 105p30 105p30 PR8-X PR8-X PR8-X
[0206] Reassortant influenza viruses which contain a backbone
according to number 3, 4, 10, 11, 14 and 16-20 are rescuable.
Influenza viruses which contain backbones number 3, 4, 10, 11 or 16
achieve viral titres of less than 10.sup.2 IU/mL. Influenza viruses
containing backbone numbers 17 and 18 achieve viral titres between
10.sup.2 and 10.sup.6 IU/mL and influenza viruses having backbone
numbers 19 and 20 even achieve titres of more than 10.sup.6
IU/mL.
[0207] These data show that influenza viruses in which the PB 1 and
PB2 segments come from the same influenza donor strain can show a
higher rescue efficiency compared with influenza viruses in which
these segments come from different influenza donor strains.
Growth Characteristics of Reassortant Influenza Viruses Containing
Backbone Segments from Two Donor Strains
[0208] Reassortant influenza strains are created which contain
backbone numbers 17, 18, 19 and 20 (as shown in table 1 above) and
the HA and NA segments from a H3 influenza strain (strain 1). As
controls, the equivalent wildtype H3 influenza virus, and a
reassortant influenza virus comprising the same HA and NA segments
and all backbone segments from PR8-X are used.
[0209] Furthermore, reassortant influenza strains are produced
which contain backbone numbers 17 and 19 and the HA and NA segments
from either a second H3 influenza (strain 1) virus or a pandemic H1
influenza virus (strain 3). As controls for the H3 strain, the
equivalent wildtype H3 (strain 2) influenza virus, and a
reassortant influenza virus comprising the same HA and NA segments
and all backbone segments from PR8-X is used. For the pandemic H1
influenza virus a reassortant influenza virus comprising the same
HA and NA segments and all backbone segments from PR8-X is
used.
[0210] The reassortant influenza viruses and the control viruses
are grown in MDCK cells and the viral titre is measured by FFA at
different time points. For the reassortant H3 viruses (strain 1)
containing backbones 17, 19 and 20, and the H3 influenza viruses
(strain 3) containing backbones 17 and 19, the influenza viruses
containing backbone segments from two donor strains grow to higher
titres compared with the wildtype virus and the reassortant virus
which contains backbone segments from only a single donor strain
(see FIG. 13, FIG. 14 and FIG. 15(A)).
[0211] For the pandemic H1 influenza virus, the reassortant
influenza strains containing backbones 17 and 19 grow to higher
titres compared with the control which contained all backbone
segments from PR8-X (see FIG. 9).
[0212] The data show that reassortant influenza viruses which
contain backbone segments from two different donor strains can show
improved growth rates compared with reassortant influenza viruses
which contain backbone segments from only a single donor
strain.
[0213] The experiments were also repeated using reassortant
influenza viruses which contain backbone 19 or the backbone
segments from PR8-X in combination with the HA and NA segments from
four different H1 strains or a H3 strain. The results are shown in
FIG. 10, FIG. 11, and FIG. 12.
Reassortant Influenza Viruses with Backbone Segments from Two
Different Donor Strains Give Higher Yields
[0214] To test whether reassortant influenza viruses containing
backbone segments from two different influenza donor strains can
also provide higher yields, the HA yield of the reassortant strains
is tested by HA-ELISA. To this end, the same reassortant influenza
viruses as described above containing backbone #19 and the HA/NA
segments of the H3 (strain 2) and H1 influenza strains are used. As
controls, the equivalent wildtype influenza viruses and reassortant
influenza viruses comprising the same HA and NA segments and all
backbone segments from PR8-X are used. In addition, the viral
titres are confirmed with a FFA assay.
[0215] The results confirm that the reassortant influenza strains
which contain backbone segments from two different donor strains
can grow to higher yields compared with influenza viruses which
contained all backbones from PR8-X (see FIG. 16 (A) and FIG. 17
(C)). Furthermore, reassortant influenza viruses comprising
backbone segments from two donor strains also give higher HA yields
(see FIG. 16 (B) and FIG. 17 (D)).
[0216] These data show that reassortant influenza viruses which
contain backbone segments from two donor strains give higher yields
compared with reassortant influenza viruses which contain backbone
segments from only a single donor strains.
[0217] It will be understood that the invention has been described
by way of example only and modifications may be made whilst
remaining within the scope and spirit of the invention.
TABLE-US-00002 SEQUENCES SEQUENCE: 1 (PA, A/New Caledonia/20/1999)
GATTCGAAATGGAAGATTTTGTGCGACAATGCTTCAATCCGATGATTGTCGAGCTTGCGGAAAAGGCAATGAAA-
G
AGTATGGAGAGGACCTGAAAATCGAAACAAACAAATTTGCAGCAATATGCACTCACTTGGAAGTATGCTTCATG-
T
ATTCAGATTTTCATTTCATCAATGAGCAAGGCGAATCAATAATAGTAGAGCCTGAGGACCCAAATGCACTTTTA-
A
AGCACAGATTTGAGATAATAGAGGGACGAGATCGTACAATGGCATGGACAGTTGTAAACAGTATTTGCAACACC-
A
CAGGAGCTGAGAAACCAAAGTTTCTGCCAGATCTGTATGATTACAAAGAGAATAGATTCATCGAGATTGGAGTG-
A
CAAGGAGGGAAGTTCACATATACTATCTGGAAAAGGCCAACAAAATTAAATCTGAGAAGACACACATTCACATT-
T
TCTCATTCACTGGCGAAGAAATGGCCACAAAGGCCGATTACACTCTCGATGAAGAAAGCAGGGCTAGGATTAAA-
A
CCAGACTATTCACCATAAGACAAGAAATGGCAAGCAGAGGTCTTTGGGACTCCTTTCGTCAGTCCGAAAGAGGC-
G
AAGAAACAATTGAAGAAAGATTTGAAATCACAGGGACAATGCGCAGGCTCGCTGACCAAAGCCTTCCGCCGAAC-
T
TCTCCTGCATTGAGAATTTTAGAGCCTATGTGGATGGATTTGAACCGAACGGCTACATTGAGGGCAAGCTTTCT-
C
AAATGTCCAAAGAAGTAAATGCTAGAATTGAGCCTTTTTTGAAAACAACACCACGACCAATTAGACTTCCGGAT-
G
GGCCTCCTTGTTTTCAGCGGTCAAAATTCCTGCTGATGGATTCTTTAAAATTAAGCATTGAGGATCCAAATCAT-
G
AAGGAGAGGGAATACCACTATATGATGCAATCAAGTGTATGAGAACATTCTTTGGATGGAAAGAACCCTCTGTT-
G
TCAAGCCACACGGGAAGGGAATAAATCCGAATTATCTGCTGTCATGGAAGCAGGTATTGGAAGAGCTGCAGGAC-
A
TTGAGAGTGAGGAGAAGATTCCAAGAACAAAAAACATGAAAAAAACGAGTCAGCTAAAGTGGGCACTTGGTGAG-
A
ACATGGCACCAGAGAAGGTGGATTTTGATGACTGTAAAGATATAAGCGATTTGAAGCAATATGATAGTGACGAA-
C
CTGAATTAAGGTCATTTTCAAGTTGGATCCAGAATGAGTTCAACAAGGCATGCGAGCTGACCGATTCAATCTGG-
A
TAGAGCTCGATGAGATTGGAGAAGATGTGGCCCCGATTGAACACATTGCAAGCATGAGAAGAAATTACTTCACA-
G
CTGAGGTGTCCCATTGCAGAGCCACAGAATATATAATGAAGGGGGTATACATTAATACTGCTTTGCTTAATGCA-
T
CCTGTGCAGCAATGGATGATTTCCAACTAATTCCCATGATAAGCAAATGTAGAACTAAAGAGGGAAGGAGAAAG-
A
CCAATTTGTACGGCTTCATCGTAAAAGGAAGATCTCACTTAAGGAATGACACCGATGTGGTAAACTTTGTGAGC-
A
TGGAGTTTTCCCTCACTGACCCAAGACTTGAGCCACACAAATGGGAGAAGTACTGTGTTCTTGAGATAGGAGAT-
A
TGCTTCTAAGGAGTGCAATAGGCCAAGTGTCAAGGCCCATGTTCTTGTATGTAAGGACAAATGGAACCTCAAAA-
A
TTAAAATGAAATGGGGAATGGAGATGAGGCGTTGCCTCCTCCAATCCCTTCAACAAATAGAGAGCATGATTGAA-
G
CTGAGTCCTCCGTCAAGGAGAAAGACATGACAAAAGAGTTTTTTGAGAATAGATCAGAAACATGGCCCATTGGA-
G
AGTCACCAAAAGGAGTGGAAGAAGGTTCCATTGGGAAAGTATGCAGGACACTATTGGCTAAGTCAGTATTCAAT-
A
GTCTGTATGCATCTCCACAATTAGAAGGATTTTCAGCTGAGTCAAGAAAGTTGCTCCTCATTGTTCAGGCTCTT-
A
GGGACAATCTGGAACCTGGGACCTTTGATCTTGGGGGGCTATATGAAGCAATTGAGGAGTGCCTGATTAATGAT-
C
CCTGGGTTTTGCTTAATGCTTCTTGGTTCAACTCCTTCCTAACACATGCATTGAGATAGCTGGGGCAATGCTAC-
T ATTTACTATCCATACTGTCCAAAAAA SEQUENCE: 2 (PB1, A/New
Caledonia/20/1999)
AATGGATGTCAATCCGACATTACTTTTCTTAAAAGTGCCAGCACAAAATGCTATAAGCACAACTTTTCCTTATA-
C
TGGTGACCCTCCTTACAGCCATGGGACAGGAACAGGGTACACCATGGATACAGTCAACAGGACACATCAGTACT-
C
AGAAAGAGGAAGATGGACAAAAAATACCGAAACTGGAGCACCGCAACTCAACCCAATTGATGGGCCACTACCAA-
A
AGACAATGAACCAAGTGGCTATGCCCAAACAGATTGTGTATTAGAAGCAATGGCTTTCCTTGAGGAATCCCATC-
C
TGGTATTTTTGAAAACTCTTGTATTGAAACAATGGAGGTTGTTCAGCAAACAAGGGTGGACAAACTGACACAAG-
G
CAGACAGACCTATGACTGGACTCTAAATAGGAACCAGCCTGCTGCCACAGCATTGGCCAACACTATAGAAGTGT-
T
CAGATCAAACGGCCTCATAGCAAATGAATCTGGGAGGCTAATAGACTTCCTTAAAGATGTAATGGAGTCGATGG-
A
CAGAGACGAAGTAGAGATCACAACTCATTTTCAAAGAAAGAGGAGAGTGAGAGACAATGTAACTAAAAAAATGG-
T
GACCCAAAGAACAATAGGCAAAAAGAAACATAAATTAGACAAAAGAAGTTACCTAATTAGGGCATTAACCCTGA-
A
CACAATGACCAAAGATGCTGAGAGGGGGAAACTAAAACGCAGAGCAATTGCAACCCCAGGAATGCAAATAAGGG-
G
GTTTGTATACTTTGTTGAGACACTGGCAAGAAGCATATGTGAAAAGCTTGAACAATCAGGGTTGCCAGTTGGAG-
G
AAATGAAAAGAAAGCAAAGTTAGCAAATGTTGTAAGGAAGATGATGACCAACTCCCAGGACACTGAAATTTCTT-
T
CACCATCACTGGAGATAACACAAAATGGAACGAAAATCAAAACCCTAGAATGTTCTTGGCCATGATCACATATA-
T
AACCAAAAATCAGCCTGAATGGTTCAGAAATATTCTAAGTATTGCTCCAATAATGTTTTCAAACAAAATGGCGA-
G
ACTAGGTAAGGGGTACATGTTTGAAAGCAAGAGTATGAAACTGAGAACTCAAATACCTGCAGAGATGCTAGCCA-
A
CATAGATTTGAAATATTTCAATGATTCAACTAAAAAGAAAATTGAAAAAATCCGGCCATTATTAATAGATGGAA-
C
TGCATCATTGAGTCCTGGAATGATGATGGGCATGTTCAATATGTTAAGCACCGTCTTGGGCGTCTCCATTCTGA-
A
TCTTGGGCAAAAGAGATACACCAAGACTACTTACTGGTGGGATGGTCTTCAATCGTCTGATGATTTTGCTCTGA-
T
TGTGAATGCACCCAACTATGCAGGAATTCAAGCTGGAGTTGACAGGTTTTATCGAACCTGTAAGCTGCTCGGAA-
T
TAATATGAGCAAAAAGAAGTCTTACATAAACAGAACAGGTACCTTTGAGTTCACGAGCTTTTTCTATCGTTATG-
G
GTTTGTTGCCAATTTCAGCATGGAGCTTCCTAGTTTTGGGGTGTCTGGGGTCAATGAATCTGCAGACATGAGTA-
T
TGGAGTCACTGTCATCAAAAACAATATGATAAACAATGACCTTGGCCCAGCAACTGCTCAAATGGCCCTTCAGT-
T
ATTTATAAAAGATTACAGGTACACGTATCGATGCCACAGAGGTGACACACAAATACAAACCCGGAGATCATTTG-
A
GATAAAGAAACTATGGGACCAAACCCGCTCCAAAGCTGGGCTGTTGGTCTCTGATGGAGGCCCCAATTTATATA-
A
CATTAGAAATCTCCATATTCCTGAAGTCTGCTTGAAATGGGAGTTGATGGATGAGGATTACCAGGGGCGTTTAT-
G
CAACCCATTGAACCCGTTTGTCAGTCATAAAGAGATTGAATCAGTGAACAATGCAGTGATGATGCCGGCACATG-
G
TCCAGCCAAAAATATGGAGTATGACGCTGTTGCAACAACACACTCCTGGGTTCCCAAAAGGAATCGATCCATTT-
T
GAATACGAGCCAAAGGGGGATACTTGAGGATGAGCAAATGTATCAGAGGTGCTGCAATTTATTTGAAAAATTCT-
T
CCCAAGTAGCTCATACAGAAGACCAGTTGGAATATCCAGTATGGTAGAGGCTATGGTTTCCAGAGCCCGAATTG-
A
TGCACGGATTGATTTCGAATCTGGAAGGATAAAAAAAGAGGAATTCGCTGAGATCATGAAGACCTGTTCCACCA-
T TGAAGACCTCAGACGGCAAAAATAGGGAATTTGGCTTGTCCTTCATGAAAA SEQUENCE: 3
(PB2, A/New Caledonia/20/1999)
AATATGGAAAGAATAAAAGAGCTAAGGAATCTGATGTCACAATCTCGCACTCGCGAGATACTTACAAAAACTAC-
T
GTAGACCACATGGCCATAATCAAGAAATACACATCAGGAAGACAGGAGAAAAACCCATCACTTAGAATGAAATG-
G
ATGATGGCAATGAAATACCCAATTACAGCAGATAAAAGGATAACGGAAATGATTCCTGAAAGAAATGAGCAAGG-
A
CAGACATTATGGAGTAAAGTGAATGATGCCGGATCAGACCGAGTGATGATATCACCCCTGGCTGTGACATGGTG-
G
AACAGAAATGGACCAGTGGCAAGTACTATTCACTATCCAAAAATCTACAAAACTTACTTTGAAAAGGTTGAAAG-
G
TTAAAACATGGAACCTTTGGCCCTGTACACTTTAGAAACCAAGTCAAAATACGCCGAAGAGTCGACATAAATCC-
T
GGTCATGCAGACCTCAGCGCCAAGGAGGCACAGGATGTAATTATGGAAGTTGTTTTCCCTAATGAAGTGGGAGC-
C
AGAATACTAACATCAGAATCGCAATTAACGATAACCAAGGAGAAAAAAGAAGAACTCCAGAATTGCAAAATTTC-
C
CCTTTGATGGTTGCATACATGTTAGAGAGGGAACTTGTCCGCAAAACGAGATTTCTCCCGGTTGCTGGTGGAAC-
A
AGCAGTGTGTACATTGAAGTTTTGCATTTAACACAGGGGACATGCTGGGAGCAGATGTACACTCCAGGTGGGGA-
G
GTGAGGAATGATGATGTTGATCAAAGCCTAATTATTGCTGCTAGGAACATAGTGAGAAGAGCTGCAGTATCAGC-
A
GATCCACTAGCATCTTTATTAGAAATGTGCCATAGCACACAGATTGGTGGGACAAGGATGGTGGATATTCTCAG-
G
CAAAATCCAACAGAAGAACAAGCTGTGGATATATGCAAAGCAGCAATGGGGCTGAGAATCAGTTCATCCTTCAG-
T
TTTGGCGGATTCACATTTAAGAGAACAAGTGGATCATCAGTCAAAAGGGAGGAAGAAGTGCTCACGGGCAATCT-
G
CAAACATTGAAGCTAACTGTGCATGAGGGATATGAAGAGTTCACAATGGTTGGGAAAAGGGCAACAGCTATACT-
C
AGAAAAGCAACCAGGAGATTGATTCAACTAATAGTGAGTGGAAGAGACGAACAGTCAATAGTCGAAGCAATAGT-
T
GTAGCAATGGTATTCTCACAAGAAGATTGCATGGTAAAAGCAGTTAGAGGTGATCTGAATTTCGTTAATAGAGC-
G
AATCAGCGGTTGAATCCCATGCATCAACTTTTGAGACATTTTCAGAAGGATGCTAAAGTACTTTTCTTAAATTG-
G
GGAATTGAACCTATCGACAATGTGATGGGAATGATTGGGATATTACCTGATATGACTCCAAGTACCGAGATGTC-
A
ATGAGAGGAGTGAGAGTCAGCAAAATGGGTGTAGATGAATACTCCAATGCTGAAAGGGTAGTGGTGAGCATTGA-
C
CGTTTTTTGAGAGTCCGGGACCAAAGAGGAAATGTACTACTGTCTCCAGAGGAAGTCAGTGAAACACAGGGAAC-
A
GAGAAACTGACAATAACTTACTCTTCATCAATGATGTGGGAGATTAATGGCCCTGAGTCAGTGTTGATCAATAC-
C
TATCAGTGGATCATCAGAAACTGGGAGACTGTTAAAATTCAGTGGTCTCAGAACCCTACAATGCTATACAATAA-
A
ATGGAATTCGAGCCATTTCAGTCTCTAGTCCCTAAGGCCATTAGAGGCCAATACAGTGGGTTTGTTAGAACTCT-
A
TTTCAACAAATGAGGGATGTGCTTGGGACCTTTGACACAACTCAGATAATAAAACTTCTTCCCTTTGCAGCCGC-
T
CCACCAAAGCAAAGTAGAATGCAATTCTCATCATTGACTGTGAATGTGAGGGGATCAGGAATGAGAATACTTGT-
A
AGGGGTAATTCTCCAGTATTCAACTACAACAAGACCACTAAGAGACTCACAGTCCTCGGAAAGGATGCTGGCAC-
T
TTAACTGAAGACCCAGATGAAGGCACAGCTGGAGTGGAATCTGCTGTTCTAAGGGGATTCCTCATTCTAGGCAA-
A
GAAGATAGAAGATATGGGCCAGCATTAAGCATCAATGAATTGAGCAACCTTGCGAAAGGGGAAAAAGCTAATGT-
G
CTAATTGGGCAAGGGGACGTAGTGTTGGTAATGAAACGAAAACGGGACTCTAGCATACTTACTGACAGCCAGAC-
A GCGACCAAAAGAATTCGGATGGCCATCAATTAATTTCGAATAATTTAAA SEQUENCE: 4
(NP, A/New Caledonia/20/1999)
ATCACTCACTGAGTGACATCAAAGTCATGGCGTCCCAAGGCACCAAACGGTCTTACGAACAGATGGAGACTGAT-
G
GGGAACGCCAGAATGCAACTGAAATCAGAGCATCCGTCGGAAGAATGATTGGTGGAATTGGGCGATTCTACATC-
C
AAATGTGCACCGAGCTTAAACTCAATGATTATGAGGGACGACTGATCCAGAACAGCTTGACAATAGAGAGAATG-
G
TGCTCTCTGCTTTTGATGAGAGGAGGAATAAATATCTGGAAGAACATCCCAGCGCGGGGAAAGATCCTAAGAAA-
A
CTGGAGGACCCATATACAAGAGAGTAGATGGAAAGTGGGTGAGGGAACTCGTCCTTTATGACAAAGAAGAAATA-
A
GGCGGATTTGGCGCCAAGCCAACAATGGTGATGATGCAACGGCTGGTTTGACTCACATTATGATCTGGCATTCT-
A
ATTTGAATGATACAACTTACCAGAGGACAAGAGCTCTTGTCCGCACCGGAATGGATCCCAGGATGTGCTCTTTG-
A
TGCAAGGTTCAACTCTCCCTAGAAGATCTGGAGCAGCAGGCGCTGCAGTCAAAGGAGTTGGGACAATGGTGTTG-
G
AGTTAATCAGGATGATCAAACGTGGGATCAATGACCGAAACTTCTGGAGGGGTGAGAATGGAAGAAAAACAAGG-
A
TTGCTTATGAGAGAATGTGCAACATTCTCAAAGGAAAATTTCAAACAGCTGCACAAAAAGCAATGATGGATCAA-
G
TGAGAGAAAGCCGGAACCCAGGAAATGCTGAGATCGAAGATCTCACTTTTCTGGCACGGTCTGCACTCATATTA-
A
GAGGGTCAGTTGCTCACAAGTCTTGCCTGCCTGCCTGTGTGTATGGACCAGCCGTAGCCAGTGGGTACGACTTC-
G
AAAAAGAGGGATACTCTTTGGTAGGGGTAGACCCTTTTAAACTGCTTCAAACCAGTCAGGTATACAGCCTAATC-
A
GACCAAACGAGAATCCCGCACACAAGAGTCAGTTGGTGTGGATGGCATGCAATTCTGCTGCATTTGAAGATCTA-
A
GAGTGTCAAGCTTCATCAGAGGGACAAGAGTACTTCCAAGGGGGAAGCTCTCCACTAGAGGAGTACAAATTGCT-
T
CAAATGAAAACATGGATGCTATTGTATCAAGTACTCTTGAACTGAGAAGCAGATACTGGGCCATAAGAACCAGA-
A
GTGGAGGGAACACTAATCAACAAAGGGCCTCTGCGGGCCAAATCAGCACACAACCTACGTTTTCTGTGCAGAGA-
A
ACCTCCCATTTGACAAAACAACCATCATGGCAGCATTCACTGGGAATACGGAGGGAAGAACATCAGACATGAGG-
G
CAGAAATCATAAAGATGATGGAAAGTGCAAGACCAGAAGAAGTGTCCTTCCAGGGGCGGGGAGTCTTTGAGCTC-
T
CGGACGAAAGGGCAACGAACCCGATCGTGCCCTCCTTTGACATGAGTAATGAAGGATCTTATTTCTTCGGAGAC-
A ATGCAGAGGAGTACGACAATTAATGAA SEQUENCE: 5 (M, A/New
Caledonia/20/1999)
GATGAGTCTTCTAACCGAGGTCGAAACGTACGTTCTCTCTATCGTCCCGTCAGGCCCCCTCAAAGCCGAGATCG-
C
ACAGAGACTTGAAAATGTCTTTGCTGGAAAGAATACCGATCTTGAGGCTCTCATGGAATGGCTAAAGACAAGAC-
C
AATCCTGTCACCTCTGACTAAGGGGATTTTAGGATTTGTGTTCACGCTCACCGTGCCCAGTGAGCGAGGACTGC-
A
GCGTAGACGCTTTGTCCAAAATGCCCTTAATGGGAATGGGGATCCAAATAATATGGACAGAGCAGTTAAACTGT-
A
TCGAAAGCTTAAGAGGGAGATAACATTCCATGGGGCCAAAGAAATAGCACTCAGTTATTCTGCTGGTGCACTTG-
C
CAGTTGTATGGGACTCATATACAACAGGATGGGGGCTGTGACCACCGAATCAGCATTTGGCCTTATATGCGCAA-
C
CTGTGAACAGATTGCCGACTCCCAGCATAAGTCTCATAGGCAAATGGTAACAACAACCAACCCATTAATAAGAC-
A
TGAGAACAGAATGGTTCTGGCCAGCACTACAGCTAAGGCTATGGAGCAAATGGCTGGATCGAGTGAACAAGCAG-
C
TGAGGCCATGGAGGTTGCTAGTCAGGCCAGGCAGATGGTGCAGGCAATGAGAGCCATTGGGACTCATCCTAGCT-
C
TAGCACTGGTCTGAAAAATGATCTCCTTGAAAATTTGCAGGCCTATCAGAAACGAATGGGGGTGCAGATGCAAC-
G
ATTCAAGTGATCCTCTTGTTGTTGCCGCAAGTATAATTGGGATTGTGCACCTGATATTGTGGATTATTGATCGC-
C
TTTTTTCCAAAAGCATTTATCGTATCTTTAAACACGGTTTAAAAAGAGGGCCTTCTACGGAAGGAGTACCAGAG-
T
CTATGAGGGAAGAATATCGAGAGGAACAGCAGAATGCTGTGGATGCTGACGATGGTCATTTTGTCAGCATAGAG-
C TAGAGTAAA SEQUENCE: 6 (NS, A/New Caledonia/20/1999)
ATGGATTCCCACACTGTGTCAAGCTTTCAGGTAGATTGCTTCCTTTGGCATGTCCGCAAACAAGTTGCAGACCA-
A
GATCTAGGCGATGCCCCATTCCTTGATCGGCTTCGCCGAGATCAGAAGTCTCTAAAGGGAAGAGGCAGCACTCT-
C
GGTCTGAACATCGAAACAGCCACTTGTGTTGGAAAGCAAATAGTAGAGAGGATTCTGAAAGAAGAATCCGATGA-
G
GCATTTAAAATGACCATGGCCTCCGCACTTGCTTCGCGGTACCTAACTGACATGACTATTGAAGAAATGTCAAG-
G
GACTGGTTCATGCTCATGCCCAAGCAGAAAGTGGCTGGCCCTCTTTGTGTCAGAATGGACCAGGCGATAATGGA-
T
AAGAACATCATACTGAAAGCGAATTTCAGTGTGATTTTTGACCGGTTGGAGAATCTGACATTACTAAGGGCTTT-
C
ACCGAAGAGGGAGCAATTGTTGGCGAAATTTCACCATTGCCTTCTCTTCCAGGACATACTAATGAGGATGTCAA-
A
AATGCAATTGGGGTCCTCATCGGGGGACTTGAATGGAATGATAACACAGTTCGAGTCTCTGAAACTCTACAGAG-
A
TTCGCTTGGAGAAGCAGTAATGAGACTGGGGGACCTCCATTCACTCCAACACAGAAACGGAAAATGGCGGGAAC-
A
ATTAGGTCAGAAGTTTGAAGAAATAAGATGGCTGATTGAAGAAGTGAGGCATAAATTGAAGACGACAGAGAATA-
G
TTTTGAGCAAATAACATTTATGCAAGCATTACAGCTATTGTTTGAAGTGGAACAAGAGATTAGAACGTTTTCGT-
T TCAGCTTATTTAATGATAA SEQUENCE: 7 (HA, A/New Caledonia/20/1999)
CCAAAATGAAAGCAAAACTACTGGTCCTGTTATGTACATTTACAGCTACATATGCAGACACAATATGTATAGGC-
T
ACCATGCCAACAACTCAACCGACACTGTTGACACAGTACTTGAGAAGAATGTGACAGTGACACACTCTGTCAAC-
C
TACTTGAGGACAGTCACAATGGAAAACTATGTCTACTAAAAGGAATAGCCCCACTACAATTGGGTAATTGCAGC-
G
TTGCCGGATGGATCTTAGGAAACCCAGAATGCGAATTACTGATTTCCAAGGAATCATGGTCCTACATTGTAGAA-
A
CACCAAATCCTGAGAATGGAACATGTTACCCAGGGTATTTCGCCGACTATGAGGAACTGAGGGAGCAATTGAGT-
T
CAGTATCTTCATTTGAGAGATTCGAAATATTCCCCAAAGAAAGCTCATGGCCCAACCACACCGTAACCGGAGTA-
T
CAGCATCATGCTCCCATAATGGGAAAAGCAGTTTTTACAGAAATTTGCTATGGCTGACGGGGAAGAATGGTTTG-
T
ACCCAAACCTGAGCAAGTCCTATGTAAACAACAAAGAGAAAGAAGTCCTTGTACTATGGGGTGTTCATCACCCG-
C
CTAACATAGGGAACCAAAGGGCCCTCTATCATACAGAAAATGCTTATGTCTCTGTAGTGTCTTCACATTATAGC-
A
GAAGATTCACCCCAGAAATAGCCAAAAGACCCAAAGTAAGAGATCAGGAAGGAAGAATCAACTACTACTGGACT-
C
TGCTGGAACCTGGGGATACAATAATATTTGAGGCAAATGGAAATCTAATAGCGCCATGGTATGCTTTTGCACTG-
A
GTAGAGGCTTTGGATCAGGAATCATCACCTCAAATGCACCAATGGATGAATGTGATGCGAAGTGTCAAACACCT-
C
AGGGAGCTATAAACAGCAGTCTTCCTTTCCAGAATGTACACCCAGTCACAATAGGAGAGTGTCCAAAGTATGTC-
A
GGAGTGCAAAATTAAGGATGGTTACAGGACTAAGGAACATCCCATCCATTCAATCCAGAGGTTTGTTTGGAGCC-
A
TTGCCGGTTTCATTGAAGGGGGGTGGACTGGAATGGTAGATGGGTGGTATGGTTATCATCATCAGAATGAGCAA-
G
GATCTGGCTATGCTGCAGATCAAAAAAGTACACAAAATGCCATTAACGGGATTACAAACAAGGTGAATTCTGTA-
A
TTGAGAAAATGAACACTCAATTCACAGCTGTGGGCAAAGAATTCAACAAATTGGAAAGAAGGATGGAAAACTTA-
A
ATAAAAAAGTTGATGATGGGTTTCTAGACATTTGGACATATAATGCAGAATTGTTGGTTCTACTGGAAAATGAA-
A
GGACTTTGGATTTCCATGACTCCAATGTGAAGAATCTGTATGAGAAAGTAAAAAGCCAATTAAAGAATAATGCC-
A
AAGAAATAGGAAACGGGTGTTTTGAATTCTATCACAAGTGTAACAATGAATGCATGGAGAGTGTGAAAAATGGA-
A
CTTATGACTATCCAAAATATTCCGAAGAATCAAAGTTAAACAGGGAGAAAATTGATGGAGTGAAATTGGAATCA-
A
TGGGAGTCTATCAGATTCTGGCGATCTACTCAACTGTCGCCAGTTCCCTGGTTCTTTTGGTCTCCCTGGGGGCA-
A
TCAGCTTCTGGATGTGTTCCAATGGGTCTTTGCAGTGTAGAATATGCATCTGAGACCAGAATTTCAGAAATATA-
A GAA SEQUENCE: 8 (NA, A/New Caledonia/20/1999)
AATGAATCCAAATCAAAAAATAATAACCATTGGATCAATCAGTATAGCAATCGGAATAATTAGTCTAATGTTGC-
A
AATAGGAAATATTATTTCAATATGGGCTAGTCACTCAATCCAAACTGGAAGTCAAAACCACACTGGAGTATGCA-
A
CCAAAGAATCATCACATATGAAAACAGCACCTGGGTGAATCACACATATGTTAATATTAACAACACTAATGTTG-
T
TGCTGGAAAGGACAAAACTTCAGTGACATTGGCCGGCAATTCATCTCTTTGTTCTATCAGTGGATGGGCTATAT-
A
CACAAAAGACAACAGCATAAGAATTGGCTCCAAAGGAGATGTTTTTGTCATAAGAGAACCTTTCATATCATGTT-
C
TCACTTGGAATGCAGAACCTTTTTTCTGACCCAAGGTGCTCTATTAAATGACAAACATTCAAATGGGACCGTTA-
A
GGACAGAAGTCCTTATAGGGCCTTAATGAGCTGTCCTCTAGGTGAAGCTCCGTCCCCATACAATTCAAAGTTTG-
A
ATCAGTTGCATGGTCAGCAAGCGCATGCCATGATGGCATGGGCTGGTTAACAATCGGAATTTCTGGTCCAGACA-
A
TGGAGCTGTGGCTGTACTAAAATACAACGGCATAATAACTGAAACCATAAAAAGTTGGAAAAAGCGAATATTAA-
G
AACACAAGAGTCTGAATGTGTCTGTGTGAACGGGTCATGTTTCACCATAATGACCGATGGCCCGAGTAATGGGG-
C
CGCCTCGTACAAAATCTTCAAGATCGAAAAGGGGAAGGTTACTAAATCAATAGAGTTGAATGCACCCAATTTTC-
A
TTATGAGGAATGTTCCTGTTACCCAGACACTGGCACAGTGATGTGTGTATGCAGGGACAACTGGCATGGTTCAA-
A
TCGACCTTGGGTGTCTTTTAATCAAAACCTGGATTATCAAATAGGATACATCTGCAGTGGGGTGTTCGGTGACA-
A
TCCGCGTCCCAAAGATGGAGAGGGCAGCTGTAATCCAGTGACTGTTGATGGAGCAGACGGAGTAAAGGGGTTTT-
C
ATACAAATATGGTAATGGTGTTTGGATAGGAAGGACTAAAAGTAACAGACTTAGAAAGGGGTTTGAGATGATTT-
G
GGATCCTAATGGATGGACAGATACCGACAGTGATTTCTCAGTGAAACAGGATGTTGTGGCAATAACTGATTGGT-
C
AGGGTACAGCGGAAGTTTCGTTCAACATCCTGAGTTAACAGGATTGGACTGTATAAGACCTTGCTTCTGGGTTG-
A
GTTAGTCAGAGGACTGCCTAGAGAAAATACAACAATCTGGACTAGTGGGAGCAGCATTTCTTTTTGTGGCGTAA-
A TAGTGATACTGCAAACTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATTGACAAGTAG
SEQUENCE: 9 (PA, PR8-X)
AGCGAAAGCAGGTACTGATCCAAAATGGAAGATTTTGTGCGACAATGCTTCAATCCGATGATTGTCGAGCTTGC-
G
GAAAAAACAATGAAAGAGTATGGGGAGGACCTGAAAATCGAAACAAACAAATTTGCAGCAATATGCACTCACTT-
G
GAAGTATGCTTCATGTATTCAGATTTTCACTTCATCAATGAGCAAGGCGAGTCAATAATCGTAGAACTTGGTGA-
T
CCAAATGCACTTTTGAAGCACAGATTTGAAATAATCGAGGGAAGAGATCGCACAATGGCCTGGACAGTAGTAAA-
C
AGTATTTGCAACACTACAGGGGCTGAGAAACCAAAGTTTCTACCAGATTTGTATGATTACAAGGAGAATAGATT-
T
ATCGAAATTGGAGTAACAAGGAGAGAAGTTCACATATACTATCTGGAAAAGGCCAATAAAATTAAATCTGAGAA-
A
ACACACATCCACATTTTCTCGTTCACTGGGGAAGAAATGGCCACAAAGGCAGACTACACTCTCGATGAAGAAAG-
C
AGGGCTAGGATCAAAACCAGACTATTCACCATAAGACAAGAAATGGCCAGCAGAGGCCTCTGGGATTCCTTTCG-
T
CAGTCCGAGAGAGGAGAAGAGACAATTGAAGAAAGGTTTGAAATCACAGGAACAATGCGCAAGCTTGCCGACCA-
A
AGTCTCCCGCCGAACTTCTCCAGCCTTGAAAATTTTAGAGCCTATGTGGATGGATTCGAACCGAACGGCTACAT-
T
GAGGGCAAGCTGTCTCAAATGTCCAAAGAAGTAAATGCTAGAATTGAACCTTTTTTGAAAACAACACCACGACC-
A
CTTAGACTTCCGAATGGGCCTCCCTGTTCTCAGCGGTCCAAATTCCTGCTGATGGATGCCTTAAAATTAAGCAT-
T
GAGGACCCAAGTCATGAAGGAGAGGGAATACCGCTATATGATGCAATCAAATGCATGAGAACATTCTTTGGATG-
G
AAGGAACCCAATGTTGTTAAACCACACGAAAAGGGAATAAATCCAAATTATCTTCTGTCATGGAAGCAAGTACT-
G
GCAGAACTGCAGGACATTGAGAATGAGGAGAAAATTCCAAAGACTAAAAATATGAAGAAAACAAGTCAGCTAAA-
G
TGGGCACTTGGTGAGAACATGGCACCAGAAAAGGTAGACTTTGACGACTGTAAAGATGTAGGTGATTTGAAGCA-
A
TATGATAGTGATGAACCAGAATTGAGGTCGCTTGCAAGTTGGATTCAGAATGAGTTTAACAAGGCATGCGAACT-
G
ACAGATTCAAGCTGGATAGAGCTCGATGAGATTGGAGAAGATGTGGCTCCAATTGAACACATTGCAAGCATGAG-
A
AGGAATTATTTCACATCAGAGGTGTCTCACTGCAGAGCCACAGAATACATAATGAAGGGGGTGTACATCAATAC-
T
GCCTTGCTTAATGCATCTTGTGCAGCAATGGATGATTTCCAATTAATTCCAATGATAAGCAAGTGTAGAACTAA-
G
GAGGGAAGGCGAAAGACCAACTTGTATGGTTTCATCATAAAAGGAAGATCCCACTTAAGGAATGACACCGACGT-
G
GTAAACTTTGTGAGCATGGAGTTTTCTCTCACTGACCCAAGACTTGAACCACATAAATGGGAGAAGTACTGTGT-
T
CTTGAGATAGGAGATATGCTTATAAGAAGTGCCATAGGCCAGGTTTCAAGGCCCATGTTCTTGTATGTGAGAAC-
A
AATGGAACCTCAAAAATTAAAATGAAATGGGGAATGGAGATGAGGCGTTGCCTCCTCCAGTCACTTCAACAAAT-
T
GAGAGTATGATTGAAGCTGAGTCCTCTGTCAAAGAGAAAGACATGACCAAAGAGTTCTTTGAGAACAAATCAGA-
A
ACATGGCCCATTGGAGAGTCCCCCAAAGGAGTGGAGGAAAGTTCCATTGGGAAGGTCTGCAGGACTTTATTAGC-
A
AAGTCGGTATTCAACAGCTTGTATGCATCTCCACAACTAGAAGGATTTTCAGCTGAATCAAGAAAACTGCTTCT-
T
ATCGTTCAGGCTCTTAGGGACAACCTTGAACCTGGGACCTTTGATCTTGGGGGGCTATATGAAGCAATTGAGGA-
G
TGCCTGATTAATGATCCCTGGGTTTTGCTTAATGCTTCTTGGTTCAACTCCTTCCTTACACATGCATTGAGTTA-
G TTGTGGCAGTGCTACTATTTGCTATCCATACTGTCCAAAAAAGTACCTTGTTTCTACT
SEQUENCE: 10 (PB1, PR8-X)
AGCGAAAGCAGGCAAACCATTTGAATGGATGTCAATCCGACCTTACTTTTCTTAAAAGTGCCAACACAAAATGC-
T
ATAAGCACAACTTTCCCTTATACTGGAGACCCTCCTTACAGCCATGGGACAGGAACAGGATACACCATGGATAC-
T
GTCAACAGGACACATCAGTACTCAGAAAAGGGAAGATGGACAACAAACACCGAAACTGGAGCACCGCAACTCAA-
C
CCGATTGATGGGCCACTGCCAGAAGACAATGAACCAAGTGGTTATGCCCAAACAGATTGTGTATTGGAGGCGAT-
G
GCTTTCCTTGAGGAATCCCATCCTGGTATTTTTGAAAACTCGTGTATTGAAACGATGGAGGTTGTTCAGCAAAC-
A
CGAGTAGACAAGCTGACACAAGGCCGACAGACCTATGACTGGACTCTAAATAGAAACCAACCTGCTGCAACAGC-
A
TTGGCCAACACAATAGAAGTGTTCAGATCAAATGGCCTCACGGCCAATGAGTCTGGAAGGCTCATAGACTTCCT-
T
AAGGATGTAATGGAGTCAATGAACAAAGAAGAAATGGGGATCACAACTCATTTTCAGAGAAAGAGACGGGTGAG-
A
GACAATATGACTAAGAAAATGATAACACAGAGAACAATGGGTAAAAAGAAGCAGAGATTGAACAAAAGGAGTTA-
T
CTAATTAGAGCATTGACCCTGAACACAATGACCAAAGATGCTGAGAGAGGGAAGCTAAAACGGAGAGCAATTGC-
A
ACCCCAGGGATGCAAATAAGGGGGTTTGTATACTTTGTTGAGACACTGGCAAGGAGTATATGTGAGAAACTTGA-
A
CAATCAGGGTTGCCAGTTGGAGGCAATGAGAAGAAAGCAAAGTTGGCAAATGTTGTAAGGAAGATGATGACCAA-
T
TCTCAGGACACCGAACTTTCTTTCACCATCACTGGAGATAACACCAAATGGAACGAAAATCAGAATCCTCGGAT-
G
TTTTTGGCCATGATCACATATATGACCAGAAATCAGCCCGAATGGTTCAGAAATGTTCTAAGTATTGCTCCAAT-
A
ATGTTCTCAAACAAAATGGCGAGACTGGGAAAAGGGTATATGTTTGAGAGCAAGAGTATGAAACTTAGAACTCA-
A
ATACCTGCAGAAATGCTAGCAAGCATCGATTTGAAATATTTCAATGATTCAACAAGAAAGAAGATTGAAAAAAT-
C
CGACCGCTCTTAATAGAGGGGACTGCATCATTGAGCCCTGGAATGATGATGGGCATGTTCAATATGTTAAGCAC-
T
GTATTAGGCGTCTCCATCCTGAATCTTGGACAAAAGAGATACACCAAGACTACTTACTGGTGGGATGGTCTTCA-
A
TCCTCTGACGATTTTGCTCTGATTGTGAATGCACCCAATCATGAAGGGATTCAAGCCGGAGTCGACAGGTTTTA-
T
CGAACCTGTAAGCTACTTGGAATCAATATGAGCAAGAAAAAGTCTTACATAAACAGAACAGGTACATTTGAATT-
C
ACAAGTTTTTTCTATCGTTATGGGTTTGTTGCCAATTTCAGCATGGAGCTTCCCAGTTTTGGGGTGTCTGGGAT-
C
AACGAGTCAGCGGACATGAGTATTGGAGTTACTGTCATCAAAAACAATATGATAAACAATGATCTTGGTCCAGC-
A
ACAGCTCAAATGGCCCTTCAGTTGTTCATCAAAGATTACAGGTACACGTACCGATGCCATAGAGGTGACACACA-
A
ATACAAACCCGAAGATCATTTGAAATAAAGAAACTGTGGGAGCAAACCCGTTCCAAAGCTGGACTGCTGGTCTC-
C
GACGGAGGCCCAAATTTATACAACATTAGAAATCTCCACATTCCTGAAGTCTGCCTAAAATGGGAATTGATGGA-
T
GAGGATTACCAGGGGCGTTTATGCAACCCACTGAACCCATTTGTCAGCCATAAAGAAATTGAATCAATGAACAA-
T
GCAGTGATGATGCCAGCACATGGTCCAGCCAAAAACATGGAGTATGATGCTGTTGCAACAACACACTCCTGGAT-
C
CCCAAAAGAAATCGATCCATCTTGAATACAAGTCAAAGAGGAGTACTTGAGGATGAACAAATGTACCAAAGGTG-
C
TGCAATTTATTTGAAAAATTCTTCCCCAGCAGTTCATACAGAAGACCAGTCGGGATATCCAGTATGGTGGAGGC-
T
ATGGTTTCCAGAGCCCGAATTGATGCACGGATTGATTTCGAATCTGGAAGGATAAAGAAAGAAGAGTTCACTGA-
G
ATCATGAAGATCTGTTCCACCATTGAAGAGCTCAGACGGCAAAAATAGTGAATTTAGCTTGTCCTTCATGAAAA-
A ATGCCTTGTTTCTACT SEQUENCE: 11 (PB2, PR8-X)
AGCGAAAGCAGGTCAATTATATTCAATATGGAAAGAATAAAAGAACTAAGAAATCTAATGTCGCAGTCTCGCAC-
C
CGCGAGATACTCACAAAAACCACCGTGGACCATATGGCCATAATCAAGAAGTACACATCAGGAAGACAGGAGAA-
G
AACCCAGCACTTAGGATGAAATGGATGATGGCAATGAAATATCCAATTACAGCAGACAAGAGGATAACGGAAAT-
G
ATTCCTGAGAGAAATGAGCAAGGACAAACTTTATGGAGTAAAATGAATGATGCCGGATCAGACCGAGTGATGGT-
A
TCACCTCTGGCTGTGACATGGTGGAATAGGAATGGACCAATAACAAATACAGTTCATTATCCAAAAATCTACAA-
A
ACTTATTTTGAAAGAGTAGAAAGGCTAAAGCATGGAACCTTTGGCCCTGTCCATTTTAGAAACCAAGTCAAAAT-
A
CGTCGGAGAGTTGACATAAATCCTGGTCATGCAGATCTCAGTGCCAAGGAGGCACAGGATGTAATCATGGAAGT-
T
GTTTTCCCTAACGAAGTGGGAGCCAGGATACTAACATCGGAATCGCAACTAACGATAACCAAAGAGAAGAAAGA-
A
GAACTCCAGGATTGCAAAATTTCTCCTTTGATGGTTGCATACATGTTGGAGAGAGAACTGGTCCGCAAAACGAG-
A
TTCCTCCCAGTGGCTGGTGGAACAAGCAGTGTGTACATTGAAGTGTTGCATTTGACTCAAGGAACATGCTGGGA-
A
CAGATGTATACTCCAGGAGGGGAAGTGAGGAATGATGATGTTGATCAAAGCTTGATTATTGCTGCTAGGAACAT-
A
GTGAGAAGAGCTGCAGTATCAGCAGATCCACTAGCATCTTTATTGGAGATGTGCCACAGCACACAGATTGGTGG-
A
ATTAGGATGGTAGACATCCTTAGGCAGAACCCAACAGAAGAGCAAGCCGTGGATATATGCAAGGCTGCAATGGG-
A
CTGAGAATTAGCTCATCCTTCAGTTTTGGTGGATTCACATTTAAGAGAACAAGCGGATCATCAGTCAAGAGAGA-
G
GAAGAGGTGCTTACGGGAAATCTTCAAACATTGAAGATAAGAGTGCATGAGGGATATGAAGAGTTCACAATGGT-
T
GGGAGAAGAGCAACAGCCATACTCAGAAAAGCAACCAGGAGATTGATTCAGCTGATAGTGAGTGGGAGAGACGA-
A
CAGTCGATTGCCGAAGCAATAATTGTGGCCATGGTATTTTCACAAGAGGATTGTATGATAAAAGCAGTCAGAGG-
T
GATCTGAATTTCGTCAATAGGGCGAATCAGCGATTGAATCCTATGCATCAACTTTTAAGACATTTTCAGAAGGA-
T
GCGAGAGTGCTTTTTCAAAATTGGGGAGTTGAACCTATCGACAATGTGATGGGAATGATTGGGATATTGCCCGA-
C
ATGACTCCAAGCATCGAGATGTCAATGAGAGGAGTGAGAATCAGCAAAATGGGTGTAGATGAGTACTCCAGCAC-
G
GAGAGGGTAGTGGTGAGCATTGACCGTTTTTTGAGAATCCGGGACCAACGAGGAAATGTACTACTGTCTCCCGA-
G
GAGGTCAGTGAAACACAGGGAACAGAGAAACTGACAATAACTTACTCATCGTCAATGATGTGGGAGATTAATGG-
T
CCTGAATCAGTATTGGTCAATACCTATCAATGGATCATCAGAAACTGGGAAACTGTTAAAATTCAGTGGTCCCA-
G
AACCCTACAATGCTATACAATAAAATGGAATTTGAACCATTTCAGTCTTTAGTACCTAAGGCCATTAGAGGCCA-
A
TACAGTGGGTTTGTAAGAACTCTGTTCCAACAAATGAGGGATGTGCTTGGGACATTTGATACCGCACAGATAAT-
A
AAACTTCTTCCCTTCGCAGCCGCTCCACCAAAGCAAAGTAGAATGCAGTTCTCCTCATTTACTGTGAATGTGAG-
G
GGATCAGGAATGAGAATACTTGTAAGGGGCAATTCTCCTGTATTCAACTATAACAAGGCCACGAAGAGACTCAC-
A
GTTCTCGGAAAGGATGCTGGCACTTTAACTGAAGACCCAGATGAAGGCACAGCTGGAGTGGAGTCCGCTGTTCT-
G
AGGGGATTCCTCATTCTGGGCAAAGAAGACAAGAGATATGGGCCAGCACTAAGCATCAATGAACTGAGCAACCT-
T
GCGAAAGGAGAGAAGGCTAATGTGCTAATTGGGCAAGGAGACGTGGTGTTGGTAATGAAACGGAAACGGGACTC-
T
AGCATACTTACTGACAGCCAGACAGCGACCAAAAGAATTCGGATGGCCATCAATTAGTGTCGAATAGTTTAAAA-
A CGACCTTGTTTCTACT SEQUENCE: 12 (NP, PR8-X)
AGCAAAAGCAGGGTAGATAATCACTCACTGAGTGACATCAAAATCATGGCGTCTCAAGGCACCAAACGATCTTA-
C
GAACAGATGGAGACTGATGGAGAACGCCAGAATGCCACTGAAATCAGAGCATCCGTCGGAAAAATGATTGGTGG-
A
ATTGGACGATTCTACATCCAAATGTGCACCGAACTCAAACTCAGTGATTATGAGGGACGGTTGATCCAAAACAG-
C
TTAACAATAGAGAGAATGGTGCTCTCTGCTTTTGACGAAAGGAGAAATAAATACCTTGAAGAACATCCCAGTGC-
G
GGAAAAGATCCTAAGAAAACTGGAGGACCTATATACAGGAGAGTAAACGGAAAGTGGATGAGAGAACTCATCCT-
T
TATGACAAAGAAGAAATAAGGCGAATCTGGCGCCAAGCTAATAATGGTGACGATGCAACGGCTGGTCTGACTCA-
C
ATGATGATCTGGCATTCCAATTTGAATGATGCAACTTATCAGAGGACAAGAGCTCTTGTTCGCACCGGAATGGA-
T
CCCAGGATGTGCTCTCTGATGCAAGGTTCAACTCTCCCTAGGAGGTCTGGAGCCGCAGGTGCTGCAGTCAAAGG-
A
GTTGGAACAATGGTGATGGAATTGGTCAGAATGATCAAACGTGGGATCAATGATCGGAACTTCTGGAGGGGTGA-
G
AATGGACGAAAAACAAGAATTGCTTATGAAAGAATGTGCAACATTCTCAAAGGGAAATTTCAAACTGCTGCACA-
A
AAAGCAATGATGGATCAAGTGAGAGAGAGCCGGAACCCAGGGAATGCTGAGTTCGAAGATCTCACTTTTCTAGC-
A
CGGTCTGCACTCATATTGAGAGGGTCGGTTGCTCACAAGTCCTGCCTGCCTGCCTGTGTGTATGGACCTGCCGT-
A
GCCAGTGGGTACGACTTTGAAAGGGAGGGATACTCTCTAGTCGGAATAGACCCTTTCAGACTGCTTCAAAACAG-
C
CAAGTGTACAGCCTAATCAGACCAAATGAGAATCCAGCACACAAGAGTCAACTGGTGTGGATGGCATGCCATTC-
T
GCCGCATTTGAAGATCTAAGAGTATTAAGCTTCATCAAAGGGACGAAGGTGCTCCCAAGAGGGAAGCTTTCCAC-
T
AGAGGAGTTCAAATTGCTTCCAATGAAAATATGGAGACTATGGAATCAAGTACACTTGAACTGAGAAGCAGGTA-
C
TGGGCCATAAGGACCAGAAGTGGAGGAAACACCAATCAACAGAGGGCATCTGCGGGCCAAATCAGCATACAACC-
T
ACGTTCTCAGTACAGAGAAATCTCCCTTTTGACAGAACAACCATTATGGCAGCATTCAATGGGAATACAGAGGG-
G
AGAACATCTGACATGAGGACCGAAATCATAAGGATGATGGAAAGTGCAAGACCAGAAGATGTGTCTTTCCAGGG-
G
CGGGGAGTCTTCGAGCTCTCGGACGAAAAGGCAGCGAGCCCGATCGTGCCTTCCTTTGACATGAGTAATGAAGG-
A TCTTATTTCTTCGGAGACAATGCAGAGGAGTACGACAATTAAAGAAAAATACCCTTGTTTCTACT
SEQUENCE: 13 (M, PR8-X)
AGCAAAAGCAGGTAGATATTGAAAGATGAGTCTTCTAACCGAGGTCGAAACGTACGTACTCTCTATCATCCCGT-
C
AGGCCCCCTCAAAGCCGAGATCGCACAGAGACTTGAAGATGTCTTTGCAGGGAAGAACACCGATCTTGAGGTTC-
T
CATGGAATGGCTAAAGACAAGACCAATCCTGTCACCTCTGACTAAGGGGATTTTAGGATTTGTGTTCACGCTCA-
C
CGTGCCCAGTGAGCGAGGACTGCAGCGTAGACGCTTTGTCCAAAATGCCCTTAATGGGAACGGGGATCCAAATA-
A
CATGGACAAAGCAGTTAAACTGTATAGGAAGCTCAAGAGGGAGATAACATTCCATGGGGCCAAAGAAATCTCAC-
T
CAGTTATTCTGCTGGTGCACTTGCCAGTTGTATGGGCCTCATATACAACAGGATGGGGGCTGTGACCACTGAAG-
T
GGCATTTGGCCTGGTATGTGCAACCTGTGAACAGATTGCTGACTCCCAGCATCGGTCTCATAGGCAAATGGTGA-
C
AACAACCAATCCACTAATCAGACATGAGAACAGAATGGTTTTAGCCAGCACTACAGCTAAGGCTATGGAGCAAA-
T
GGCTGGATCGAGTGAGCAAGCAGCAGAGGCCATGGAGGTTGCTAGTCAGGCTAGACAAATGGTGCAAGCGATGA-
G
AACCATTGGGACTCATCCTAGCTCCAGTGCTGGTCTGAAAAATGATCTTCTTGAAAATTTGCAGGCCTATCAGA-
A
ACGAATGGGGGTGCAGATGCAACGGTTCAAGTGATCCTCTCACTATTGCCGCAAATATCATTGGGATCTTGCAC-
T
TGACATTGTGGATTCTTGATCGTCTTTTTTTCAAATGCATTTACCGTCGCTTTAAATACGGACTGAAAGGAGGG-
C
CTTCTACGGAAGGAGTGCCAAAGTCTATGAGGGAAGAATATCGAAAGGAACAGCAGAGTGCTGTGGATGCTGAC-
G ATGGTCATTTTGTCAGCATAGAGCTGGAGTAAAAAACTACCTTGTTTCTACT SEQUENCE: 14
(NS, PR8-X)
AGCAAAAGCAGGGTGACAAAAACATAATGGATCCAAACACTGTGTCAAGCTTTCAGGTAGATTGCTTTCTTTGG-
C
ATGTCCGCAAACGAGTTGCAGACCAAGAACTAGGTGATGCCCCATTCCTTGATCGGCTTCGCCGAGATCAGAAA-
T
CCCTAAGAGGAAGGGGCAGTACTCTCGGTCTGGACATCAAGACAGCCACACGTGCTGGAAAGCAGATAGTGGAG-
C
GGATTCTGAAAGAAGAATCCGATGAGGCACTTAAAATGACCATGGCCTCTGTACCTGCGTCGCGTTACCTAACT-
G
ACATGACTCTTGAGGAAATGTCAAGGGACTGGTCCATGCTCATACCCAAGCAGAAAGTGGCAGGCCCTCTTTGT-
A
TCAGAATGGACCAGGCGATCATGGATAAGAACATCATACTGAAAGCGAACTTCAGTGTGATTTTTGACCGGCTG-
G
AGACTCTAATATTGCTAAGGGCTTTCACCGAAGAGGGAGCAATTGTTGGCGAAATTTCACCATTGCCTTCTCTT-
C
CAGGACATACTGCTGAGGATGTCAAAAATGCAGTTGGAGTCCTCATCGGAGGACTTGAATGGAATGATAACACA-
G
TTCGAGTCTCTGAAACTCTACAGAGATTCGCTTGGAGAAGCAGTAATGAGAATGGGAGACCTCCACTCACTCCA-
A
AACAGAAACGAGAAATGGCGGGAACAATTAGGTCAGAAGTTTGAAGAAATAAGATGGTTGATTGAAGAAGTGAG-
A
CACAAACTGAAGATAACAGAGAATAGTTTTGAGCAAATAACATTTATGCAAGCCTTACATCTATTGCTTGAAGT-
G GAGCAAGAGATAAGAACTTTCTCGTTTCAGCTTATTTAGTACTAAAAAACACCCTTGTTTCTACT
SEQUENCE: 15 (HA, PR8-X)
AGCAAAAGCAGGGGAAAATAAAAACAACCAAAATGAAGGCAAACCTACTGGTCCTGTTATGTGCACTTGCAGCT-
G
CAGATGCAGACACAATATGTATAGGCTACCATACGAACAATTCAACCGACACTGTTGACACAGTACTCGAGAAG-
A
ATGTGACAGTGACACACTCTGTTAACCTGCTCGAAGACAGCCACAACGGAAAACTATGTAGATTAAAAGGAATA-
G
CCCCACTACAATTGGGGAAATGTAACATCGCCGGATGGCTCTTGGGAAACCCAGAATGCGACCCACTGCTTCCA-
G
TGAGATCATGGTCCTACATTGTAGAAACACCAAACTCTGAGAATGGAATATGTTATCCAGGAGATTTCATCGAC-
T
ATGAGGAGCTGAGGGAGCAATTGAGCTCAGTGTCATCATTCGAAAGATTCGAAATATTTCCCAAAGAAAGCTCA-
T
GGCCCAACCACAACACAAACGGAGTAACGGCAGCATGCTCCCATGAGGGGAAAAGCAGTTTTTACAGAAATTTG-
C
TATGGCTGACGGAGAAGGAGGGCTCATACCCAAAGCTGAAAAATTCTTATGTGAACAAAAAAGGGAAAGAAGTC-
C
TTGTACTGTGGGGTATTCATCACCCGCCTAACAGTAAGGAACAACAGAATCTCTATCAGAATGAAAATGCTTAT-
G
TCTCTGTAGTGACTTCAAATTATAACAGGAGATTTACCCCGGAAATAGCAGAAAGACCCAAAGTAAGAGATCAA-
G
CTGGGAGGATGAACTATTACTGGACCTTGCTAAAACCCGGAGACACAATAATATTTGAGGCAAATGGAAATCTA-
A
TAGCACCAATGTATGCTTTCGCACTGAGTAGAGGCTTTGGGTCCGGCATCATCACCTCAAACGCATCAATGCAT-
G
AGTGTAACACGAAGTGTCAAACACCCCTGGGAGCTATAAACAGCAGTCTCCCTTACCAGAATATACACCCAGTC-
A
CAATAGGAGAGTGCCCAAAATACGTCAGGAGTGCCAAATTGAGGATGGTTACAGGACTAAGGAACATTCCGTCC-
A
TTCAATCCAGAGGTCTATTTGGAGCCATTGCCGGTTTTATTGAAGGGGGATGGACTGGAATGATAGATGGATGG-
T
ATGGTTATCATCATCAGAATGAACAGGGATCAGGCTATGCAGCGGATCAAAAAAGCACACAAAATGCCATTAAC-
G
GGATTACAAACAAGGTGAACACTGTTATCGAGAAAATGAACATTCAATTCACAGCTGTGGGTAAAGAATTCAAC-
A
AATTAGAAAAAAGGATGGAAAATTTAAATAAAAAAGTTGATGATGGATTTCTGGACATTTGGACATATAATGCA-
G
AATTGTTAGTTCTACTGGAAAATGAAAGGACTCTGGAATTCCATGACTCAAATGTGAAGAATCTGTATGAGAAA-
G
TAAAAAGCCAATTAAAGAATAATGCCAAAGAAATCGGAAATGGATGTTTTGAGTTCTACCACAAGTGTGACAAT-
G
AATGCATGGAAAGTGTAAGAAATGGGACTTATGATTATCCCAAATATTCAGAAGAGTCAAAGTTGAACAGGGAA-
A
AGGTAGATGGAGTGAAATTGGAATCAATGGGGATCTATCAGATTCTGGCGATCTACTCAACTGTCGCCAGTTCA-
C
TGGTGCTTTTGGTCTCCCTGGGGGCAATCAGTTTCTGGATGTGTTCTAATGGATCTTTGCAGTGCAGAATATGC-
A TCTGAGATTAGAATTTCAGAGATATGAGGAAAAACACCCTTGTTTCTACT SEQUENCE: 16
(NA, PR8-X)
AGCAAAAGCAGGGGTTTAAAATGAATCCAAATCAGAAAATAATAACCATTGGATCAATCTGTCTGGTAGTCGGA-
C
TAATTAGCCTAATATTGCAAATAGGGAATATAATCTCAATATGGATTAGCCATTCAATTCAAACTGGAAGTCAA-
A
ACCATACTGGAATATGCAACCAAAACATCATTACCTATAAAAATAGCACCTGGGTAAAGGACACAACTTCAGTG-
A
TATTAACCGGCAATTCATCTCTTTGTCCCATCCGTGGGTGGGCTATATACAGCAAAGACAATAGCATAAGAATT-
G
GTTCCAAAGGAGACGTTTTTGTCATAAGAGAGCCCTTTATTTCATGTTCTCACTTGGAATGCAGGACCTTTTTT-
C
TGACCCAAGGTGCCTTACTGAATGACAAGCATTCAAGTGGGACTGTTAAGGACAGAAGCCCTTATAGGGCCTTA-
A
TGAGCTGCCCTGTCGGTGAAGCTCCGTCCCCGTACAATTCAAGATTTGAATCGGTTGCTTGGTCAGCAAGTGCA-
T
GTCATGATGGCATGGGCTGGCTAACAATCGGAATTTCAGGTCCAGATAATGGAGCAGTGGCTGTATTAAAATAC-
A
ACGGCATAATAACTGAAACCATAAAAAGTTGGAGGAAGAAAATATTGAGGACACAAGAGTCTGAATGTGCCTGT-
G
TAAATGGTTCATGTTTTACTATAATGACTGATGGCCCGAGTGATGGGCTGGCCTCGTACAAAATTTTCAAGATC-
G
AAAAGGGGAAGGTTACTAAATCAATAGAGTTGAATGCACCTAATTCTCACTATGAGGAATGTTCCTGTTACCCT-
G
ATACCGACAAAGTGATGTGTGTGTGCAGAGACAATTGGCATGGTTCGAACCGGCCATGGGTGTCTTTCGATCAA-
A
ACCTGGATTATCAAATAGGATACATCTGCAGTGGGGTTTTCGGTGACAACCCGCGTCCCGAAGATGGAACAGGC-
A
GCTGTGGTCCAGTGTATGTTGATGGAGCAAACGGAGTAAAGGGATTTTCATATAGGTATGGTAATGGTGTTTGG-
A
TAGGAAGGACCAAAAGTCACAGTTCCAGACATGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGAGACT-
G
ATAGTAAGTTCTCTGTGAGGCAAGATGTTGTGGCAATGACTGATTGGTCAGGGTATAGCGGAAGTTTCGTTCAA-
C
ATCCTGAGCTGACAGGGCTAGACTGTATGAGGCCGTGCTTCTGGGTTGAATTAATCAGGGGACGACCTAAAGAA-
A
AAACAATCTGGACTAGTGCGAGCAGCATTTCTTTTTGTGGCGTGAATAGTGATACTGTAGATTGGTCTTGGCCA-
G ACGGTGCTGAGTTGCCATTCAGCATTGACAAGTAGTCTGTTCAAAAAACTCCTTGTTTCTACT
SEQUENCE: 17 (PA, 105p30)
AGCGAAAGCAGGTACTGATTCGAAATGGAAGATTTTGTGCGACAATGCTTCAATCCGATGATTGTCGAGCTTGC-
G
GAAAAGGCAATGAAAGAGTATGGAGAGGACCTGAAAATCGAAACAAACAAATTTGCAGCAATATGCACCCACTT-
G
GAAGTATGCTTCATGTATTCAGATTTTCATTTCATCAATGAGCAAGGCGAATCAATAATAGTAGAGCCTGAGGA-
C
CCAAATGCACTTTTAAAACACAGATTTGAGATAATAGAGGGGCGAGATCGTACAATGGCATGGACAGTTGTAAA-
C
AGTATTTGCAACACCACAGGAGCTGAGAAACCAAAGTTTCTGCCAGATCTGTATGATTACAAAGAGAATAGGTT-
C
ATCGAAATTGGAGTGACAAGGAGAGAAGTTCACATATACTATCTGGAAAAGGCCAACAAAATTAAATCTGAGAA-
G
ACACATATTCACATTTTCTCATTTACTGGCGAAGAAATGGCCACAAAGGCCGATTACACTCTCGATGAAGAAAG-
C
AGGGCTAGAATTAAAACCAGACTATTCACCATAAGGCAAGAAATGGCAAGCAGAGGTCTTTGGGACTCCTTTCG-
T
CAGTCCGAAAGAGGCGAAGAGACAATTGAAGAAAGGTTTGAAATCACAGGGACAATGCGCAGGCTCGCTGATCA-
A
AGCCTTCCGCCGAACTTCTCCTGCATTGAGAATTTTAGAGCCTATGTGGATGGATTTGAACCGAACGGCTACAT-
T
GAGGGCAAGCTTTCTCAAATGTCCAAAGAAGTAAATGCTAAAATTGAGCCTTTTTTGAAAACAACACCTCGACC-
A
ATTAGACTTCCGAATGGGCCTCCTTGTTTTCAGCGGTCAAAATTCCTGCTGATGGATTCTTTAAAATTAAGCAT-
T
GAGGATCCAAATCATGAAGGGGAGGGAATACCACTATATGATGCAATCAAGTGTATGAGAACATTCTTTGGATG-
G
AAAGAACCCACTGTTGTCAAGCCACACGAGAAGGGAATAAATCCGAATTATCTGCTGTCGTGGAAGCAGGTGTT-
G
GAAGAGCTGCAGGACATTGAGAGTGAGGAGAAGATTCCAAGAACAAAAAACATGAAAAAAACGAGTCAGTTAAA-
G
TGGGCACTTGGTGAGAACATGGCACCAGAGAAGGTGGATTTTGATGACTGTAAAGATATAAGCGATTTGAAGCA-
A
TATGATAGTGACGAACCTGAATTAAGGTCATTTTCAAGTTGGATCCAGAATGAGTTCAACAAGGCATGCGAGCT-
G
ACCGATTCAATCTGGATAGAGCTCGATGAGATTGGAGAAGATGTGGCCCCGATTGAACACATTGCAAGCATGAG-
A
AGAAATTACTTCACAGCTGAGGTGTCCCATTGCAGAGCCACTGAATATATAATGAAAGGGGTATACATTAATAC-
T
GCTTTGCTTAATGCATCCTGTGCAGCAATGGATGATTTCCAACTAATTCCTATGATAAGCAAATGTAGAACTAA-
A
GAGGGAAGGAGAAAGACCAATTTGTACGGCTTCATCATAAAAGGAAGATCTCACTTAAGGAATGATACCGATGT-
G
GTAAACTTTGTGAGCATGGAGTTTTCCCTCACTGACCCAAGACTTGAGCCACACAAATGGGAGAAGTACTGTGT-
T
CTTGAGATAGGAGATATGCTTCTAAGGAGTGCAATAGGCCAAGTGTCAAGGCCCATGTTCTTGTATGTAAGAAC-
A
AATGGAACCTCAAAAATTAAAATGAAATGGGGAATGGAGATGAGGCGTTGCCTCCTCCAATCCCTCCAACAAAT-
A
GAGAGCATGATTGAAGCTGAGTCCTCTGTCAAGGAGAAAGACATGACAAAAGAGTTTTTTGAGAATAGATCAGA-
A
ACATGGCCCATTGGAGAGTCACCAAAAGGAGTGGAAGAAGGTTCCATTGGGAAAGTATGCAGGACACTATTGGC-
T
AAATCAGTATTCAATAGTCTGTATGCATCTCCACAATTAGAAGGATTTTCAGCTGAGTCAAGAAAGTTGCTCCT-
T
ATTGTTCAGGCTCTTAGGGACAATCTGGAACCTGGGACCTTTGATCTTGGGGGACTATATGAAGCAATTGAGGA-
G
TGCCTGATTAATGATCCCTGGGTTTTGCTTAATGCTTCTTGGTTCAACTCCTTCCTAAAACATGCATTGAGATA-
G CTGAGGCAATGCTACTATTTGTTATCCATACTGTCCAAAAAAGTA SEQUENCE: 18 (PB1,
105p30)
AGCGAAAGCAGGCAAACCATTTGAATGGATGTCAATCCGACATTACTTTTCTTAAAAGTGCCAGCACAAAATGC-
T
ATAAGCACAACTTTTCCTTATACTGGTGACCCTCCTTACAGCCATGGAACAGGAACAGGATACACCATGGATAC-
A
GTCAACAGGACACATCAGTACTCAGAAAGAGGAAGATGGACGAAAAATACCGAAACTGGAGCACCGCAACTCAA-
C
CCAATTGATGGGCCACTACCAGAAGACAATGAACCAAGTGGCTATGCCCAAACAGATTGTGTATTAGAGGCAAT-
G
GCTTTCCTTGAAGAATCCCATCCTGGTATTTTTGAAAACTCTTGTATTGAAACAATGGAGGTTGTTCAGCAAAC-
A
AGGGTGGACAAACTGACACAAGGCAGACAAACCTATGACTGGACTCTAAATAGGAACCAGCCTGCTGCCACAGC-
A
TTGGCAAACACCATAGAAGTATTCAGATCAAATGGCCTCATAGCAAATGAATCTGGAAGGCTAATAGACTTCCT-
T
AAAGATGTAATGGAGTCGATGGACAGAGACGAAGTAGAGGTCACAACTCATTTTCAAAGAAAGAGGAGAGTGAG-
A
GACAATGTAACTAAAAAAATGGTGACCCAAAGAACAATAGGAAAAAAGAAACATAAATTAGACAAAAGAAGTTA-
C
CTAATTAGGGCATTAACCCTGAACACAATGACCAAAGATGCTGAGAGGGGGAAACTAAAACGCAGAGCAATTGC-
A
ACCCCAGGAATGCAAATAAGGGGGTTTGTATACTTTGTTGAGACACTGGCAAGAAGCATATGTGAAAAGCTTGA-
A
CAATCAGGGTTGCCAGTTGGAGGAAATGAGAAGAAAGCAAAGTTAGCAAATGTTGTAAGGAAGATGATGACCAA-
C
TCCCAGGACACTGAAATTTCTTTTACCATCACTGGAGATAACACAAAATGGAACGAAAATCAAAACCCTAGAAT-
G
TTCTTGGCCATGATCACATATATAACCAAAGATCAGCCTGAATGGTTCAGAAATATTCTAAGTATTGCTCCAAT-
A
ATGTTTTCAAACAAAATGGCGAGACTAGGTAGGGGGTATATGTTTGAAAGCAAGAGTATGAAACTGAGAACCCA-
A
ATACCTGCAGAGATGCTAGCCAACATAGATTTGAAATATTTCAATGATTCAACTAAAAAGAAAATTGAAAAAAT-
T
CGACCATTATTAATAGATGGAACTGCATCATTGAGTCCTGGAATGATGATGGGCATGTTCAATATGTTAAGCAC-
C
GTCTTGGGCGTTTCCATTCTGAATCTTGGGCAAAAAAGATACACCAAGACTACTTACTGGTGGGATGGTCTTCA-
A
TCGTCTGATGATTTTGCTTTGATTGTGAATGCACCCAATTATGCAGGAATTCAAGCTGGAGTTGACAGGTTTTA-
T
CGAACCTGTAAGCTGCTCGGAATTAATATGAGCAAAAAGAAGTCTTACATAAACAGAACAGGTACCTTTGAATT-
C
ACGAGCTTTTTCTATCGTTATGGGTTTGTTGCCAATTTCAGCATGGAGCTTCCTAGTTTTGGGGTGTCTGGGGT-
C
AATGAATCTGCAGACATGAGTATTGGAGTCACTGTCATCAAAAACAATATGATAAACAATGACCTTGGCCCAGC-
A
ACTGCTCAAATGGCCCTTCAGTTATTTATAAAAGATTACAGGTACACTTATCGATGCCACAGAGGTGACACACA-
A
ATACAAACCCGGAGATCATTTGAAATAAAGAAACTATGGGACCAAACCCGCTCCAAAGCTGGGCTGTTGGTCTC-
T
GATGGAGGCCCCAATTTATATAACATTAGGAATCTACATATTCCTGAAGTCTGCTTGAAATGGGAGTTGATGGA-
T
GAGGATTACCAGGGGCGTTTATGCAACCCATTGAACCCGTTTGTCAGCCATAAAGAGATTGAATCAGTGAACAA-
T
GCAGTGATAATGCCGGCACATGGTCCAGCCAAAAATATGGAGTATGACGCTGTTGCAACAACACACTCTTGGGT-
C
CCCAAAAGAAATCGATCCATTTTAAACACGAGCCAAAGAGGGATACTTGAAGATGAGCAAATGTACCAAAGGTG-
C
TGCAATTTATTTGAAAAATTCTTCCCAAGTAGCTCATACAGAAGACCAGTTGGAATATCCAGTATGGTAGAGGC-
T
ATGGTTTCAAGAGCCCGAATTGATGCACGGATTGATTTCGAATCTGGAAGGATAAAGAAAGAGGAATTCGCTGA-
G
ATCATGAAGACCTGTTCCACCATTGAAGACCTCAGACGGCAAAAATAGGGAATTTGGCTTGTCCTTCATGAAAA-
A ATGCCTTGTTTCTACT SEQUENCE: 19 (PB2, 105p30)
AGCGAAAGCAGGTCAATTATATTCAATATGGAAAGAATAAAAGAGCTAAGGAATCTGATGTCACAATCTCGCAC-
T
CGCGAGATACTTACCAAAACTACTGTAGACCACATGGCCATAATAAAGAAATACACATCAGGAAGACAGGAGAA-
A
AACCCATCACTTAGGATGAAATGGATGATGGCAATGAAATACCCAATTACAGCTGATAAAAGGATAACGGAAAT-
G
ATTCCTGAAAGAAATGAGCAAGGACAGACACTATGGAGTAAAGTGAATGATGCCGGATCAGACCGAGTGATGAT-
A
TCACCCCTAGCTGTGACATGGTGGAACAGAAATGGACCAGTGGCAAACACTATCCACTATCCAAAAATCTACAA-
A
ACTTACTTTGAAAAGGTTGAAAGGTTAAAACATGGAACCTTTGGCCCTGTACACTTTAGAAACCAAGTCAAAAT-
A
CGCCGAAGAGTCGACATAAATCCTGGTCATGCAGACCTCAGCGCCAAGGAGGCACAGGATGTAATTATGGAAGT-
T
GTTTTCCCTAATGAAGTGGGAGCCAGAATACTAACATCAGAATCGCAATTAACGATAACTAAGGAGAAAAAAGA-
G
GAACTCCAGAATTGCAAAATTTCCCCTTTGATGGTTGCATACATGTTAGAGAGGGAACTTGTCCGCAAAACAAG-
A
TTTCTCCCGGTTGCAGGTGGAACAAGCAGTGTGTACATTGAAGTTTTGCATTTAACACAGGGGACATGCTGGGA-
G
CAGATGTACACTCCAGGTGGGGAGGTGAGGAATGATGATGTTGATCAAAGCCTAATTATTGCTGCTAGGAACAT-
A
GTGAGAAGAGCTGCAGTATCAGCAGATCCACTAGCATCTTTATTAGAAATGTGCCATAGCACACAGATTGGTGG-
A
ACAAGGATGGTGGATATTCTCAGGCAAAATCCAACAGAAGAACAAGCTGTGGACATATGCAAAGCAGCAATGGG-
G
CTGAGAATCAGTTCATCCTTCAGTTTTGGCGGATTCACATTTAAGAGAACAAGTGGATCGTCAGTCAAAAGGGA-
G
GAAGAAGTGCTAACGGGCAATCTGCAAACATTGAAGCTAACTGTGCATGAGGGATATGAAGAATTCACAATAGT-
T
GGGAAAAAGGCAACAGCTATACTCAGAAAAGCAACCAGGAGATTGATTCAACTAATAGTGAGTGGAAGAGACGA-
A
CAGTCAATAGTCGAAGCAATAGTTGTAGCAATGGTATTCTCACAAGAAGATTGCATGGTAAAAGCGGTTAGAGG-
T
GATCTGAATTTCGTTAATAGAGCGAATCAGCGGTTGAATCCCATGCATCAACTTTTGAGACATTTTCAGAAGGA-
T
GCTAAAGTACTTTTCCTAAATTGGGGAATTGAACATATTGACAATGTGATGGGAATGATTGGGATATTACCTGA-
T
ATGACTCCAAGTACCGAGATGTCAATGAGAGGAGTGAGAGTCAGCAAAATGGGTGTAGATGAATACTCCAATGC-
T
GAAAGGGTAGTGGTAAGCATTGACCGTTTTTTGAGGGTCCGGGACCAAAGAGGAAATGTATTACTGTCTCCAGA-
G
GAAGTCAGTGAAACACAAGGAACAGAGAAACTGACAATAACTTACTCTTCATCATTGATGTGGGAGATTAATGG-
C
CCTGAGTCAGTGTTGATCAATACCTACCAATGGATCATCAGAAACTGGGAGACTGTTAAAATTCAGTGGTCTCA-
G
AACCCTACAATGCTATACAATAAAATGGAATTTGAGCCATTTCAATCTCTAGTCCCCAAGGCCATTAGAGGCCA-
A
TACAGTGGGTTTGTTAGAACTCTATTTCAACAAATGAGGGATGTGCTCGGGACCTTTGACACAACTCAGATAAT-
A
AAACTTCTTCCCTTTGCAGCCGCTCCACCAAAGCAAAGTAGAATGCAATTCTCGTCATTAACTGTGAATGTGAG-
G
GGATCAGGAATGAGAATACTTGTAAGGGGTAATTCTCCAGTATTCAACTACAACAAGACCACTAAGAGACTCAC-
A
ATCCTCGGAAAGGATGCTGGCACTTTAACTGAAGACCCAGATGAAGGCACAGCTGGAGTGGAATCTGCTGTTTT-
A
AGGGGATTCCTCATTCTAGGCAAAGAAGATAGAAGATATGGGCCAGCATTAAGCATCAGTGAATTGAGCAACCT-
T
GCGAAAGGGGAGAAAGCTAATGTGCTAATTGGGCAAGGGGATGTAGTGTTGGTAATGAAACGAAAACGGGACTC-
T
AGCATACTTACTGACAGCCAGACAGCGACCAAAAGAATTCGGATGGCCATCAATTAATTTCGAATAATTTAAAA-
A CGACCTTGTTTCTACT SEQUENCE: 20 (NP, 105p30)
AGCAAAAGCAGGGTAGATAATCACTCACTGAGTGACATCAAAGTCATGGCGTCCCAAGGCACCAAACGGTCTTA-
C
GAACAGATGGAGACTGATGGGGAACGCCAGAATGCAACTGAAATCAGAGCATCCGTCGGAAGAATGATTGGGGG-
A
ATTGGGCGATTCTACATCCAAATGTGCACCGAGCTTAAGCTCAATGATTATGAGGGACGACTGATCCAGAACAG-
C
TTAACAATAGAGAGAATGGTGCTTTCTGCTTTTGATGAGAGGAGAAATAAATATCTGGAAGAACATCCCAGCGC-
A
GGGAAAGATCCTAAGAAAACTGGAGGACCCATATACAAGAGAGTAGATGGAAAGTGGGTGAGGGAACTCGTCCT-
T
TATGACAAAGAAGAAATAAGGCGGATTTGGCGCCAAGCCAACAATGGTGATGATGCAACAGCTGGTTTGACTCA-
C
ATTATGATCTGGCATTCTAATTTGAATGATACAACTTACCAGAGGACAAGAGCTCTTGTCCGCACCGGAATGGA-
T
CCCAGGATGTGCTCTTTGATGCAAGGTTCAACTCTCCCTAGAAGATCTGGAGCAGCAGGCGCTGCAGTCAAAGG-
A
GTTGGGACAATGGTATTGGAGTTAATCAGGATGATCAAACGTGGGATCAACGACCGAAACTTCTGGAGGGGTGA-
G
AATGGGAGAAAAACAAGGATTGCTTATGAGAGAATGTGCAACATTCTCAAAGGAAAATTTCAAACAGCTGCACA-
A
AAAGCAATGATGGATCAAGTGAGAGAAAGCCGGAACCCAGGAAATGCTGAGATCGAAGATCTCACTTTTCTGGC-
A
CGGTCTGCACTCATATTGAGAGGATCAGTTGCTCACAAGTCTTGCCTGCCTGCTTGTGTGTATGGACCAGCCGT-
A
GCCAGTGGGTATGACTTCGAAAAAGAGGGATACTCTTTGGTGGGAGTAGACCCTTTCAAACTGCTTCAAACCAG-
T
CAGGTATACAGCCTAATTAGACCAAACGAGAATCCCGCACACAAGAGCCAGTTGGTGTGGATGGCATGCAATTC-
T
GCTGCATTTGAAGATCTAAGAGTGTCAAGCTTCATCAGAGGGACAAGAGTACTTCCAAGGGGGAAGCTCTCCAC-
T
AGAGGAGTACAAATTGCTTCAAATGAAAACATGGATGCTATTGTCTCAAGTACTCTTGAACTGAGAAGCAGATA-
C
TGGGCCATAAGAACCAGAAGTGGAGGGAACACCAATCAACAAAGGGCCTCTGCGGGCCAAATCAGCACACAACC-
T
ACGTTTTCTGTGCAGAGAAACCTCCCATTTGACAAAACAACCATCATGGCAGCATTCACTGGGAATACAGAGGG-
A
AGAACATCAGACATGCGGGCAGAAATCATAAAGATGATGGAAAGTGCAAGACCAGAAGAAGTGTCCTTCCAGGG-
A
CGGGGAGTCTTTGAGCTCTCGGACGAAAGGGCAACGAACCCGATCGTGCCCTCCTTTGACATGAGTAATGAAGG-
A TCTTATTTCTTCGGAGACAATGCAGAGGAGTACGACAATTAATGAAAAATACCCTTGTTTCTACT
SEQUENCE: 21 (M, 105p30)
AGCAAAAGCAGGTAGATATTGAAAGATGAGTCTTCTAACCGAGGTCGAAACGTACGTTCTCTCTATCGTCCCAT-
C
AGGCCCCCTCAAAGCCGAGATCGCACAGAGACTTGAAGATGTATTTGCTGGAAAGAATACCGATCTTGAGGCTC-
T
CATGGAATGGCTAAAGACAAGACCAATCCTGTCACCTCTGACTAAGGGGATTTTAGGATTTGTGTTCACGCTCA-
C
CGTGCCCAGTGAGCGAGGACTGCAGCGTAGACGCTTTGTCCAAAATGCCCTTAATGGGAATGGGGATCCAAATA-
A
TATGGACAAGGCTGTCAAACTGTATCGAAAGCTTAAGAGGGAGATAACATTCCATGGGGCCAAAGAAATAGCAC-
T
CAGTTATTCTGCTGGAGCACTTGCCAGTTGTATGGGACTCATATACAACAGGATGGGGGCTGTGACCACCGAAT-
C
AGCATTTGGCCTTATATGTGCAACCTGTGAACAGATTGCCGACTCCCAGCATAAGTCTCATAGGCAAATGGTAA-
C
AACAACCAATCCATTAATAAGACATGAGAACAGAATGGTTCTGGCCAGCACTACAGCTAAGGCTATGGAGCAAA-
T
GGCTGGATCGAGTGAACAAGCAGCTGAGGCCATGGAGGTTGCTAGTCAGGCCAGGCAGATGGTGCAGGCAATGA-
G
AGCCATTGGGACTCATCCTAGCTCTAGCACTGGTCTGAAAAATGATCTCCTTGAAAATTTGCAGGCCTATCAGA-
A
ACGAATGGGGGTGCAGATGCAACGATTCAAGTGATCCTCTTGTTGTTGCCGCAAGTATAATTGGGATTGTGCAC-
C
TGATATTGTGGATTATTGATCGCCTTTTTTCCAAAAGCATTTATCGTATTTTTAAACACGGTTTAAAAAGAGGG-
C
CTTCTACGGAAGGAGTACCGGAGTCTATGAGGGAAGAATATCGAGAGGAACAGCAGAATGCTGTGGATGCTGAC-
G ATGGTCATTTTGTCAGCATAGAGCTAGAGTAAAAAACTACCTTGTTTCTACT SEQUENCE: 22
(NS, 105p30)
AGCAAAAGCAGGGTGGCAAAGACATAATGGATTCCCACACTGTGTCAAGCTTTCAGGTAGATTGTTTCCTTTGG-
C
ATGTCCGCAAACAAGTTGCAGACCAAGATCTAGGCGATGCCCCCTTCCTTGATCGGCTTCGCCGAGATCAGAAG-
T
CTCTAAAGGGACGAGGCAACACTCTCGGTCTGAACATCGAAACAGCCACTTGTGTTGGAAAGCAAATAGTAGAG-
A
GGATTCTGAAAGAAGAATCCGATGAGACATTTAGAATGACCATGGCCTCCGCACTTGCTTCGCGGTACCTAACT-
G
ACATGACTGTTGAAGAAATGTCAAGGGACTGGTTCATGCTCATGCCCAAGCAGAAAGTGGCTGGCCCTCTTTGT-
G
TCAGAATGGACCAGGCGATAATGGATAAGAACATCATACTGAAAGCGAACTTCAGTGTGATTTTTGACCGGTTG-
G
AGAATCTGACATTACTAAGGGCTTTCACCGAAGAGGGAGCAATTGTTGGCGAAATTTCACCATTGCCTTCTTTT-
C
CAGGACATACTAATGAGGATGTCAAAAATGCAATTGGGGTCCTCATCGGGGGACTTGAATGGAATGATAACACA-
G
TTCGAGTCTCTGAAGCTCTACAGAGATTCGCTTGGAGAAGCAGTAATGAGACTGGGGGACCTCCATTCACTACA-
A
CACAGAAACGGAAAATGGCGGGAACAATTAGGTCAGAAGTTTGAAGAAATAAGATGGCTGATTGAAGAAGTGAG-
G
CATAAATTGAAGACGACAGAGAGTAGTTTTGAACAAATAACATTTATGCAAGCATTACAGCTATTGTTTGAAGT-
G GAACAAGAGATTAGAACGTTCTCGTTTCAGCTTATTTAATGATAAAAACACCCTTGTTTCTACT
SEQUENCE: 23 (HA, 105p30)
AGCGAAAGCAGGGGAAAATAAAAGCAACCAAAATGAAAGTAAAACTACTGGTTCTGTTATGTACATTTACAGCT-
A
CATATGCAGACACAATATGTATAGGCTACCATGCCAACAACTCAACCGACACTGTTGACACAGTACTTGAGAAG-
A
ATGTAACAGTGACACACTCTGTCAACCTACTTGAGGACAGTCACAATGGAAAACTATGTCTACTAAAAGGAATA-
G
CCCCACTACAATTGGGTAATTGCAGCGTTGCCGGATGGATCTTAGGAAACCCAGAATGCGAATTACTGATTTCC-
A
AGGAATCATGGTCCTACATTGTAGAAACACCAAATCCTGAGAATGGAACATGTTACCCAGGGTATTTCGCCGAC-
T
ATGAGGAACTGAGGGAGCAATTGAGTTCAGTATCTTCATTTGAAAGGTTCGAAATATTCCCCAAAGAGAGCTCA-
T
GGCCCAACCACACCGTAACCGGAGTATCAGCATCATGCTCCCATAACGGGAAAAGCAGTTTTTACAGAAATTTG-
C
TATGGCTGACGGGGAAGAATGGTTTGTACCCAAACCTGAGCAAGTCCTATGCAAACAACAAAGAGAAAGAAGTC-
C
TTGTACTATGGGGTGTTCATCACCCGCCTAACATAGGGGACCAAAGGGCCCTCTATCATACAGAAAATGCTTAT-
G
TCTCTGTAGTGTCTTCACATTATAGCAGAAGATTCACCCCAGAAATAGCCAAAAGACCCAAGGTGAGAGACCAG-
G
AAGGAAGAATCAACTACTACTGGACTCTGCTGGAACCCGGGGATACAATAATATTTGAGGCAAATGGAAATCTA-
A
TAGCGCCAAGGTATGCTTTCGCACTGAGTAGAGGCTTGGGATCAGGAATCATCACCTCAAATGCACCAATGGAT-
G
AATGTGATGCAAAGTGTCAAACACCTCAGGGAGCTATAAACAGCAGTCTTCCTTTCCAGAATGTACACCCAGTC-
A
CAATAGGAGAGTGTCCAAAGTATGTCAGGAGTGCAAAATTAAGGATGGTTACAGGACTAAGGAACATCCCATCC-
A
TTCAATCCAGAGGTTTGTTTGGAGCAATTGCCGGTTTCATTGAAGGGGGGTGGACTGGAATGGTAGATGGTTGG-
T
ATGGTTATCATCATCAGAATGAGCAAGGATCTGGGTATGCTGCAGATCAAAAAAGCACACAAAATGCCATTAAC-
G
GGATTACAAACAAGGTGAATTCTGTAATTGAGAAAATGAACACTCAATTCACAGCTGTGGGCAAAGAATTCAAC-
A
AATTGGAAAGAAGGATGGAAAACTTAAATAAAAAAGTTGATGATGGGTTTCTAGACATTTGGACCTATAATGCA-
G
AATTGTTGGTTCTACTGGAAAATGAAAGGACTTTGGATTTCCATGACTCCAACGTGAAGAATCTGTATGAGAAA-
G
TAAAAAGCCAATTAAAGAATAATGCCAAAGAAATAGGAAACGGGTGTTTTGAATTCTATCACAAGTGTAACGAT-
G
AATGCATGGAGAGTGTGAAAAATGGAACTTATGACTATCCAAAATATTCCGAAGAATCAAAGTTAAACAGAGAG-
A
AAATTGATGGAGTGAAATTGGAATCAATGGGAGTCTATCAGATTCTGGCGATCTACTCAACAGTCGCCAGTTCC-
C
TGGTTCTTTTGGTCTCCCTGGGGGCAATCAGCTTCTGGATGTGTTCCAATGGGTCTTTGCAGTGTAGAATATGC-
A TCTAAGACCAGAATTTCAGAAATATAAGGAAAAACACCCTTGTTTCTACT SEQUENCE: 24
(NA, 105p30)
AGCAAAAGCAGGAGTTTAAAATGAATCCAAATCAAAAAATAATAACCATTGGATCAATCAGTATAGCAATCGGA-
A
TAATTAGTCTAATGTTGCAAATAGGAAATATTATTTCAATATGGGCTAGTCACTCAATCCAAACTGGAAGTCAA-
A
ACCACACTGGAATATGCAACCAAAAAATCATCACATATGAAAACAGCACCTGGGTGAATCACACATATGTTAAT-
A
TTAACAACACTAATGTTGTTGCTGGAAAGGACAAAACTTCAGTGACACTGGCCGGCAATTCATCTCTTTGTCCT-
A
TCAGTGGATGGGCTATATACACAAAAGACAACAGCATAAGAATTGGCTCCAAAGGAGATGTTTTTGTCATAAGA-
G
AACCTTTCATATCATGTTCTCACTTGGAATGCAGAACCTTTTTTCTGACCCAAGGTGCTCTATTAAATGACAAA-
C
ATTCAAATGGAACCGTTAAGGACAGAAGTCCTTATAGGGCCTTAATGAGCTGTCCTCTAGGTGAAGCCCCGTCA-
C
CATACAATTCAAAGTTTGAATCAGTTGCATGGTCAGCAAGCGCATGCCATGATGGCAAGGGCTGGTTAACAATC-
G
GAATTTCTGGTCCAGACAATGGAGCTGTGGCTGTACTAAAATACAACGGAATAATAACTGAAACCATAAAAAGT-
T
GGGAAAAGCGAATATTGAGAACACAAGAGTCTGAATGTGTTTGTGTGAACGGGTCATGTTTCACCATAATGACC-
G
ATGGCCCGAGTAATGGGGCCGCCTCGTACAAAATCTTCAAGATCGAAAAGGGGAAGGTTACTAAATCAACAGAG-
T
TGAATGCACCCAATTTTCATTATGAGGAATGTTCCTGTTACCCAGACACTGGCACAGTGATGTGTGTATGCAGG-
G
ACAACTGGCATGGTTCAAATCGACCTTGGGTATCTTTTAATCAAAACTTGGATTATCAAATAGGATACATCTGC-
A
GTGGAGTGTTCGGTGACAATCCGCGTCCCAAAGATGGGAAGGGCAGCTGTAATCCAGTGACTGTTGATGGAGCA-
G
ACGGAGTTAAGGGGTTTTCATACAAATATGGTAATGGTGTTTGGATAGGAAGGACTAAAAGTAACAGACTTAGA-
A
AGGGGTTTGAGATGATTTGGGATCCTAATGGATGGACAGATACCGACAGTGATTTCTCAGTGAAACAGGATGTT-
G
TGGCAATAACTGATTGGTCAGGGTACAGCGGAAGTTTCGTCCAACATCCTGAGTTAACAGGATTGGACTGTATA-
A
GACCTTGCTTCTGGGTTGAGTTAGTCAGAGGACTGCCTAGAGAAAATACAACAATCTGGACTAGTGGGAGCAGC-
A
TTTCTTTTTGTGGCGTTGATAGTGATACTGCAAATTGGTCTTGGCCAGACGGTGCTGAGTTGCCGTTCACCATT-
G ACAAGTAGCTCGTTGAAAAAAACTCCTTGTTTCTACT SEQUENCE: 25 (HA,
A/Chile/1/1983)
MKAKLLVLLCALSATDADTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLEDNHNGKLCKLKGIAPLQLGKCSI-
A
GWILGNPECESLFSKKSWSYIAETPNSENGTCYPGYFADYEELREQLSSVSSFERFEIFPKESSWPKHNVTKGV-
T
AACSHKGKSSFYRNLLWLTEKNGSYPNLSKSYVNNKEKEVLVLWGVHHPSNIEDQKTIYRKENAYVSVVSSHYN-
R
RFTPEIAKRPKVRNQEGRINYYWTLLEPGDTIIFEANGNLIAPWYAFALSRGFGSGIITSNASMDECDAKCQTP-
Q
GAINSSLPFQNVHPVTIGECPKYVRSTKLRMVTGLRNIPSIQSRGLFGAIAGFIEGGWTGMIDGWYGYHHQNEQ-
G
SGYAADQKSTQNAINGITNKVNSIIEKMNTQFTAVGKEENKLEKRMENLNKKVDDGFLDIWTYNAELLVLLENE-
R
TLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNNECMESVKNGTYDYPKYSEESKLNREKIDGVKLES-
M GVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI SEQUENCE: 26 (NA,
A/Chile/1/1983)
MNPNQKIITIGSICMTIGIISLILQIGNIISIWVSHSIQTGSQNHTGICNQRIITYENSTWVNQTYVNINNTNV-
V
AGKDTTSVTLAGNSSLCPIRGWAIYSKDNSIRIGSKGDVFVIREPFISCSHLECRTFFLTQGALLNDKHSNGTV-
K
DRSPYRALMSCPIGEAPSPYNSRFESVAWSASACHDGMGWLTIGISGPDDGAVAVLKYNGIITETIKSWRKRIL-
R
TQESECVCVNGSCFTIMTDGPSNGPASYRIFKIEKGKITKSIELDAPNSHYEECSCYPDTGTVMCVCRDNWHGS-
N
RPWVSFNQNLDYQIGYICSGVFGDNPRPKDGKGSCDPVTVDGADGVKGFSYRYGNGVWIGRTKSNSSRKGFEMI-
W
DPNGWTDTDSNFLVKQDVVAMTDWSGYSGSFVQHPELTGLDCMRPCFWVELVRGRPREGTTVWTSGSSISFCGV-
N SDTANWSWPDGAELPFTIDK SEQUENCE: 27 (NA, A/California/04/09)
MNPNQKIITIGSVCMTIGMANLILQIGNIISIWISHSIQLGNQNQIETCNQSVITYENNTWVNQTYVNISNTNF-
A
AGQSVVSVKLAGNSSLCPVSGWAIYSKDNSVRIGSKGDVFVIREPFISCSPLECRTFFLTQGALLNDKHSNGTI-
K
DRSPYRTLMSCPIGEVPSPYNSRFESVAWSASACHDGINWLTIGISGPDNGAVAVLKYNGIITDTIKSWRNNIL-
R
TQESECACVNGSCFTVMTDGPSNGQASYKIFRIEKGKIVKSVEMNAPNYHYEECSCYPDSSEITCVCRDNWHGS-
N
RPWVSFNQNLEYQIGYICSGIFGDNPRPNDKTGSCGPVSSNGANGVKGFSFKYGNGVWIGRTKSISSRNGFEMI-
W
DPNGWTGTDNNFSIKQDIVGINEWSGYSGSFVQHPELTGLDCIRPCFWVELIRGRPKENTIWTSGSSISFCGVN-
S DTVGWSWPDGAELPFTIDK SEQUENCE: 28 (encodes the same amino acid
sequence as SEQUENCE: 3)
ATGGAACGCATTAAAGAACTGCGCAACCTGATGAGCCAGAGCCGCACCCGCGAAATTCTGACCAAAACCACCGT-
G
GATCATATGGCGATTATTAAAAAATATACCAGCGGCCGCCAGGAAAAAAACCCGAGCCTGCGCATGAAATGGAT-
G
ATGGCGATGAAATATCCGATTACCGCGGATAAACGCATTACCGAAATGATTCCGGAACGCAACGAACAGGGCCA-
G
ACCCTGTGGAGCAAAGTGAACGATGCGGGCAGCGATCGCGTGATGATTAGCCCGCTGGCGGTGACCTGGTGGAA-
C
CGCAACGGCCCGGTGGCGAGCACCATTCATTATCCGAAAATTTATAAAACCTATTTTGAAAAAGTGGAACGCCT-
G
AAACATGGCACCTTTGGCCCGGTGCATTTTCGCAACCAGGTGAAAATTCGCCGCCGCGTGGATATTAACCCGGG-
C
CATGCGGATCTGAGCGCGAAAGAAGCGCAGGATGTGATTATGGAAGTGGTGTTTCCGAACGAAGTGGGCGCGCG-
C
ATTCTGACCAGCGAAAGCCAGCTGACCATTACCAAAGAAAAAAAAGAAGAACTGCAGAACTGCAAAATTAGCCC-
G
CTGATGGTGGCGTATATGCTGGAACGCGAACTGGTGCGCAAAACCCGCTTTCTGCCGGTGGCGGGCGGCACCAG-
C
AGCGTGTATATTGAAGTGCTGCATCTGACCCAGGGCACCTGCTGGGAACAGATGTATACCCCGGGCGGCGAAGT-
G
CGCAACGATGATGTGGATCAGAGCCTGATTATTGCGGCGCGCAACATTGTGCGCCGCGCGGCGGTGAGCGCGGA-
T
CCGCTGGCGAGCCTGCTGGAAATGTGCCATAGCACCCAGATTGGCGGCACCCGCATGGTGGATATTCTGCGCCA-
G
AACCCGACCGAAGAACAGGCGGTGGATATTTGCAAAGCGGCGATGGGCCTGCGCATTAGCAGCAGCTTTAGCTT-
T
GGCGGCTTTACCTTTAAACGCACCAGCGGCAGCAGCGTGAAACGCGAAGAAGAAGTGCTGACCGGCAACCTGCA-
G
ACCCTGAAACTGACCGTGCATGAAGGCTATGAAGAATTTACCATGGTGGGCAAACGCGCGACCGCGATTCTGCG-
C
AAAGCGACCCGCCGCCTGATTCAGCTGATTGTGAGCGGCCGCGATGAACAGAGCATTGTGGAAGCGATTGTGGT-
G
GCGATGGTGTTTAGCCAGGAAGATTGCATGGTGAAAGCGGTGCGCGGCGATCTGAACTTTGTGAACCGCGCGAA-
C
CAGCGCCTGAACCCGATGCATCAGCTGCTGCGCCATTTTCAGAAAGATGCGAAAGTGCTGTTTCTGAACTGGGG-
C
ATTGAACCGATTGATAACGTGATGGGCATGATTGGCATTCTGCCGGATATGACCCCGAGCACCGAAATGAGCAT-
G
CGCGGCGTGCGCGTGAGCAAAATGGGCGTGGATGAATATAGCAACGCGGAACGCGTGGTGGTGAGCATTGATCG-
C
TTTCTGCGCGTGCGCGATCAGCGCGGCAACGTGCTGCTGAGCCCGGAAGAAGTGAGCGAAACCCAGGGCACCGA-
A
AAACTGACCATTACCTATAGCAGCAGCATGATGTGGGAAATTAACGGCCCGGAAAGCGTGCTGATTAACACCTA-
T
CAGTGGATTATTCGCAACTGGGAAACCGTGAAAATTCAGTGGAGCCAGAACCCGACCATGCTGTATAACAAAAT-
G
GAATTTGAACCGTTTCAGAGCCTGGTGCCGAAAGCGATTCGCGGCCAGTATAGCGGCTTTGTGCGCACCCTGTT-
T
CAGCAGATGCGCGATGTGCTGGGCACCTTTGATACCACCCAGATTATTAAACTGCTGCCGTTTGCGGCGGCGCC-
G
CCGAAACAGAGCCGCATGCAGTTTAGCAGCCTGACCGTGAACGTGCGCGGCAGCGGCATGCGCATTCTGGTGCG-
C
GGCAACAGCCCGGTGTTTAACTATAACAAAACCACCAAACGCCTGACCGTGCTGGGCAAAGATGCGGGCACCCT-
G
ACCGAAGATCCGGATGAAGGCACCGCGGGCGTGGAAAGCGCGGTGCTGCGCGGCTTTCTGATTCTGGGCAAAGA-
A
GATCGCCGCTATGGCCCGGCGCTGAGCATTAACGAACTGAGCAACCTGGCGAAAGGCGAAAAAGCGAACGTGCT-
G
ATTGGCCAGGGCGATGTGGTGCTGGTGATGAAACGCAAACGCGATAGCAGCATTCTGACCGATAGCCAGACCGC-
G ACCAAACGCATTCGCATGGCGATTAAC SEQUENCE: 29 (PA, A/New
Caledonia/20/1999)
medfvrqcfnpmivelaekamkeygedpkietnkfaaicthlevcfmysdfhfidergesiivesgdpnallkh-
r
feiiegrdrimawtvvnsicnttgvekpkflpdlydykenrfieigvtrrevhiyylekankiksekthihifs-
f
tgeematkadytldeesrariktrlftirqemasrslwdsfrqsergeetieekfeitgtmrkladqslppnfp-
s
lenfrayvdgfepngciegklsqmskevnakiepflrttprplrlpdgplchqrskfllmdalklsiedpsheg-
e
giplydaikcmktffgwkepnivkphekginpnylmawkqvlaelqdieneekiprtknmkrtsqlkwalgenm-
a
pekvdfddckdvgdlkqydsdepeprslaswvqnefnkaceltdsswieldeigedvapiehiasmrrnyftae-
v
shcrateyimkgvyintallnascaamddfqlipmiskcrtkegrrktnlygfiikgrshlrndtdvvnfvsme-
f
sltdprlephkwekycvleigdmllrtaigqvsrpmflyvrtngtskikmkwgmemrrcllqslqqiesmieae-
s
svkekdmtkeffenksetwpigesprgveegsigkvcrtllaksvfnslyaspqlegfsaesrklllivqalrd-
n lepgtfdlgglyeaieeclindpwvllnaswfnsflthalk SEQUENCE: 30 (PB1,
A/New Caledonia/20/1999)
mdvnptllflkvpaqnaisttfpytgdppyshgtgtgytmdtvnrthqysergrwtkntetgapqlnpidgplp-
k
dnepsgyaqtdcvleamafleeshpgifenscietmevvqqtrvdkltqgrqtydwtlnrnqpaatalantiev-
f
rsnglianesgrlidflkdvmesmdrdevevtthfqrkrrvrdnvtkkmvtqrtigkkkhkldkrsyliraltl-
n
tmtkdaergklkrraiatpgmqirgfvyfvetlarsicekleqsglpvggnekkaklanvvrkmmtnsqdteis-
f
titgdntkwnenqnprmflamityitknqpewfrnilsiapimfsnkmarlgkgymfesksmklrtqipaemla-
n
idlkyfndstkrkiekirpllidgtaslspgmmmgmfnmlstvlgvsilnlgqkrytkttywwdglqssddfal-
i
vnapnyagiqagvdrfyrtckllginmskkksyinrtgtfeftsffyrygfvanfsmelpsfgvsgvnesadms-
i
gvtviknnminndlgpataqmalqlfikdyrytyrchrgdtqiqtrrsfeikklwdqtrskagllvsdggpnly-
n
irnlhipevclkwelmdedyqgrlcnpsnpfvshkeiesvnnavmmpahgpaknmeydavatthswvpkrnrsi-
l
ntsqrgiledeqmyqrccnlfekffpsssyrrpvgissmveamvsraridaridfesgrikkeefaeimktcst-
i edlrrqk SEQUENCE: 31 (PB2, A/New Caledonia/20/1999)
merikelrnlmsqsrtreiltkttvdhmaiikkytsgrqeknpslrmkwmmamkypitadkritemiperneqg-
q
tlwskvndagsdrvmisplavtwwnrngpvastihypkiyktyfekverlkhgtfgpvhfrnqvkirrrvdinp-
g
hadlsakeaqdvimevvfpnevgariltsesqltitkekkeelqnckisplmvaymlerelvrktrflpvaggt-
s
svyievlhltqgtcweqmytpggevrnddvdqsliiaarnivrraavsadplasllemchstqiggtrmvdilr-
q
npteeqavdickaamglrisssfsfggftfkrtsgssvkreeevltgnlqtlkltvhegyeeftmvgkratail-
r
katrrliqlivsgrdeqsiveaivvamvfsqedcmvkavrgdlnfvnranqrlnpmhqllrhfqkdakvlflnw-
g
iepidnvmgmigilpdmtpstemsmrgvrvskmgvdeysnaervvvsidrflrvrdqrgnvllspeevsetqgt-
e
kltitysssmmweingpesvlintyqwiirnwetvkiqwsqnptmlynkmefepfqslvpkairgqysgfvrtl-
f
qqmrdvlgtfdttqiikllpfaaappkqsrmqfssltvnvrgsgmrilvrgnspvfnynkttkrltvlgkdagt-
l
tedpdegtagvesavlrgflilgkedrrygpalsinelsnlakgekanvligqgdvvlvmkrkrdssiltdsqt-
a tkrirmain SEQUENCE: 32 (NP, A/New Caledonia/20/1999)
masqgtkrsyeqmetdgerqnateirasvgrmiggigrfyiqmctelklndyegrliqnsltiermvlsafder-
r
nkyleehpsagkdpkktggpiykrvdgkwvrelvlydkeeirriwrqanngddataglthimiwhsnlndttyq-
r
tralvrtgmdprmcslmqgstlprrsgaagaavkgvgtmvlelirmikrgindrnfwrgengrktriayermcn-
i
lkgkfqtaaqkammdqvresrnpgnaeiedltflarsalilrgsvahksclpacvygpavasgydfekegyslv-
g
vdpfkllqtsqvyslirpnenpahksqlvwmacnsaafedlrvssfirgtrvlprgklstrgvqiasnenmdai-
v
sstlelrsrywairtrsggntnqqrasagqistqptfsvqrnlpfdkttimaaftgntegrtsdmraeiikmme-
s arpeevsfqgrgvfelsderatnpivpsfdmsnegsyffgdnaeeydn SEQUENCE: 33
(M1, A/New Caledonia/20/1999)
mslltevetyvlsivpsgplkaeiaqrlenvfagkntdlealmewlktrpilspltkgilgfvftltvpsergl-
q
rrrfvqnalngngdpnnmdravklyrklkreitfhgakeialsysagalascmgliynrmgavttesafglica-
t
ceqiadsqhkshrqmvtttnplirhenrmvlasttakameqmagsseqaaeamevasqarqmvqamraigthps-
s stglkndllenlqayqkrmgvqmqrfk SEQUENCE: 34 (NA, A/New
Caledonia/20/1999)
mnpnqkiitigsisiaigiislmlqigniisiwashsiqtgsqnhtgvcnqriityenstwvnhtyvninntnv-
v
agkdktsvtlagnsslcsisgwaiytkdnsirigskgdvfvirepfiscshlecrtffltqgallndkhsngtv-
k
drspyralmscplgeapspynskfesvawsasachdgmgwltigisgpdngavavlkyngiitetikswkkril-
r
tqesecvcvngscftimtdgpsngaasykifkiekgkvtksielnapnfhyeecscypdtgtvmcvcrdnwhgs-
n
rpwvsfnqnldyqigyicsgvfgdnprpkdgegscnpvtvdgadgvkgfsykygngvwigrtksnrlrkgfemi-
w
dpngwtdtdsdfsvkqdvvaitdwsgysgsfvqhpeltgldcirpcfwvelvrglprenttiwtsgssisfcgv-
n sdtanwswpdgaelpftidk SEQUENCE: 35 (PA, A/Wisconsin/67/2005)
medfvrqcfnpmivelaekamkeygedlkietnkfaaicthlevcfmysdfhfineqgesivvelddpnallkh-
r
feiiegrdrtmawtvvnsicnttgagkpkflpdlydykenrfieigvtrrevhiyylekankiksenthihifs-
f
tgeematkadytldeesrariktrlftirqemanrglwdsfrqsergeetieekfeitgtmrrladqslppnfs-
c
lenfrayvdgfepngciegklsqmskevnaqiepflkttprpiklpngppcyqrskfllmdalklsiedpsheg-
e
giplydaikcmktffgwkepyivkphekginsnyllswkqvlselqdieneekiprtknmkktsqlkwalgenm-
a
pekvdfencrdisdlkqydsdepelrslsswiqnefnkaceltdsvwieldeigedvapiehiasmrrnyftae-
v
shcrateyimkgvyintallnascaamddfqlipmiskcrtkegrrktnlygfiikgrshlrndtdvvnfvsme-
f
sltdprlephkwekycvleigdmllrsaigqisrpmflyvrtngtskvkmkwgmemrrcllqslqqiesmieae-
s
svkekdmtkeffenkseawpigespkgveegsigkvcrtllaksvfnslyaspqlegfsaesrklllvvqalrd-
n lepgtfdlgglyeaieeclindpwvllnaswfnsflthalk SEQUENCE: 36 (PB1,
A/Wisconsin/67/2005)
mdvnptllflkvpaqnaisttfpytgdppyshgtgtgytmdtvnrthqysekgkwttntetgapqlnpidgplp-
e
dnepsgyaqtdcvleamafleeshpgifenscletmeavqqtrvdrltqgrqtydwtlnrnqpaatalantiev-
f
rsngltanesgrlidflkdvmesmdkeemeitthfqrkrrvrdnmtkkmvtqrtigkkkqrvnkrgyliraltl-
n
tmtkdaergklkrraiatpgmqirgfvyfvetlarsicekleqsglpvggnekkaklanvvrkmmtnsqdtels-
f
titgdntkwnenqnprmflamityitknqpewfrnilsiapimfsnkmarlgkgymfeskrmklrtqipaemla-
s
idlkyfnestrkkiekirpllidgtaslspgmmmgmfnmlstvlgvsilnlgqkkytkttywwdglqssddfal-
i
vnapnhegiqagvnrfyrtcklvginmskkksyinktgtfeftsffyrygfvanfsmelpsfgvsginesadms-
i
gvtviknnminndlgpataqmalqlfikdyrytyrchrgdtqiqtrrsfelkklwdqtqsragllvsdggpnly-
n
irnlhipevclkwelmdenyrgrlcnplnpfvshkeiesvnnavvmpahgpaksmeydavatthswipkrnrsi-
l
ntsqrgiledeqmyqkccnlfekffpsssyrrpigissmveamvsraridaridfesgrikkeefseimkicst-
i eelrrqr SEQUENCE: 37 (PB2, A/Wisconsin/67/2005)
merikelrnlmsqsrtreiltkttvdhmaiikkytsgrqeknpslrmkwmmamkypitadkritemvperneqg-
q
tlwskmsdagsdrvmvsplavtwwnrngpvtstvhypkvyktyfdkverlkhgtfgpvhfrnqvkirrrvdinp-
g
hadlsakeaqdvimevvfpnevgariltsesqltitkekkeelrdckisplmvaymlerelvrktrflpvaggt-
s
siyievlhltqgtcweqmytpggevrnddvdqsliiaarnivrraavsadplasllemchstqiggtrmvdilr-
q
npteeqavdickaamglrisssfsfggftfkrtsgssvkkeeevltgnlqtlkirvhegyeeftmvgkratail-
r
katrrlvqlivsgrdeqsiaeaiivamvfsqedcmikavrgdlnfvnranqrlnpmhqllrhfqkdakvlfqnw-
g
iehidsvmgmvgvlpdmtpstemsmrgirvskmgvdeysstervvvsidrflrvrdqrgnvllspeevsetqgt-
e
rltitysssmmweingpesvlvntyqwiirnweavkiqwsqnpamlynkmefepfqslvpkairsqysgfvrtl-
f
qqmrdvlgtfdttqiikllpfaaappkqsrmqfssltvnvrgsgmrilvrgnspvfnynkttkrltilgkdagt-
l
iedpdestsgvesavlrgfliigkedrrygpalsinelsnlakgekanvligqgdvvlvmkrkrdssiltdsqt-
a tkrirmain SEQUENCE: 38 (NP, A/Wisconsin/67/2005)
masqgtkrsyeqmetdgdrqnateirasvgkmidgigrfyiqmctelklsdyegrliqnsltiekmvlsafder-
r
nkyleehpsagkdpkktggpiyrrvdgkwmrelvlydkeeirriwrqanngedataglthimiwhsnlndatyq-
r
tralvrtgmdprmcslmqgstlprrsgaagaavkgigtmvmelirmvkrgindrnfwrgengrktrsayermcn-
i
lkgkfqtaaqramvdqvresrnpgnaeiedliflarsalilrgsvahksclpacvygpavssgynfekegyslv-
g
idpfkllqnsqvyslirpnenpahksqlvwmachsaafedlrllsfirgtkvsprgklstrgvqiasnenmdnm-
g
sgtlelrsgywairtrsggntnqqrasagqtsvqptfsvqrnlpfekstimaaftgntegrtsdmraeiirmme-
g akpeevsfrgrgvfelsdekatnpivpsfdmsnegsyffgdnaeeydn SEQUENCE: 39
(M1, A/Wisconsin/67/2005)
mslltevetyvlsivpsgplkaeiaqrledvfagkntdlealmewlktrpilspltkgilgfvftltvpsergl-
q
rrrfvqnalngngdpnnmdkavklyrklkreitfhgakeialsysagalascmgliynrmgavttevafglvca-
t
ceqiadsqhrshrqmvattnplirhenrmvlasttakameqmagsseqaaeameiasqarqmvqamraigthps-
s stglrddllenlqtyqkrmgvqmqrfk SEQUENCE: 40 (M2,
A/Wisconsin/67/2005)
mslltevetpirnewgcrcndssdplvvaaniigilhlilwildrlffkcvyrlfkhglkrgpstegvpesmre-
e yrkeqqnavdaddshfvsiele SEQUENCE: 41 (NS, A/Wisconsin/67/2005)
AATGGATTCCAACACTGTGTCAAGTTTCCAGGTAGATTGCTTTCTTTGGCATATCCGGAAACAAGTTGTAGACC-
A
AGAACTGAGTGATGCCCCATTCCTTGATCGGCTTCGCCGAGATCAGAGGTCCCTAAGGGGAAGAGGCAATACTC-
T
CGGTCTAGACATCAAAGCAGCCACCCATGTTGGAAAGCAAATTGTAGAAAAGATTCTGAAAGAAGAATCTGATG-
A
GGCACTTAAAATGACCATGGTCTCCACACCTGCTTCGCGATACATAACTGACATGACTATTGAGGAATTGTCAA-
G
AAACTGGTTCATGCTAATGCCCAAGCAGAAAGTGGAAGGACCTCTTTGCATCAGAATGGACCAGGCAATCATGG-
A
GAAAAACATCATGTTGAAAGCGAATTTCAGTGTGATTTCTGACCGACTAGAGACCATAGTATTACTAAGGGCTT-
T
CACCGAAGAGGGAGCAATTGTTGGCGAAATCTCACCATTGCCTTCTTTTCCAGGACATACTATTGAGGATGTCA-
A
AAATGCAATTGGGGTCCTCATCGGAGGACTTGAATGGAATGATAACACAGTTCGAGTCTCTAAAAATCTACAGA-
G
ATTCGCTTGGAGAAGCAGTAATGAGAATGGGGGACCTCCACTTACTCCAAAACAGAAACGGAAAATGGCGAGAA-
C
AGCTAGGTCAAAAGTTTGAAGAGATAAGATGGCTGATTGAAGAAGTGAGACACAGACTAAAAACAACTGAAAAT-
A
GCTTTGAACAAATAACATTCATGCAAGCATTACAACTGCTGTTTGAAGTGGAACAGGAGATAAGAACTTTCTCA-
T TTCAGCTTATTTAATGATAAA SEQUENCE: 42 (HA, A/Wisconsin/67/2005)
mktiialsyilclvfaqklpgndnstatlclghhavpngtivktitndqievtnatelvqssstggicdsphqi-
l
dgenctlidallgdpqcdgfqnkkwdlfverskaysncypydvpdyaslrslvassgtlefndesfnwtgvtqn-
g
tsssckrrsnnsffsrlnwlthlkfkypalnvtmpnnekfdklyiwgvhhpvtdndqiflyaqasgritvstkr-
s
qqtvipnigsrprirnipsrisiywtivkpgdillinstgnliaprgyfkirsgkssimrsdapigkcnsecit-
p
ngsipndkpfqnvnritygacpryvkqntlklatgmrnvpekqtrgifgaiagfiengwegmvdgwygfrhqns-
e
gigqaadlkstqaainqingklnrligktnekfhqiekefsevegriqdlekyvedtkidlwsynaellvalen-
q
htidltdsemnklfertkkqlrenaedmgngcfkiyhkcdnacigsirngtydhdvyrdealnnrfqikgvelk-
s gykdwilwisfaiscfllcvallgfimwacqkgnircnici SEQUENCE: 43 (NA,
A/Wisconsin/67/2005)
mnpnqkiitigsvsltisticffmqiailittvtlhfkqyefnsppnnqvmlceptiierniteivyltnttie-
k
eicpklaeyrnwskpqcnitgfapfskdnsirlsaggdiwvtrepyvscdpdkcyqfalgqgttlnnvhsndtv-
h
drtpyrtllmnelgvpfhlgtkqvciawsssschdgkawlhvcvtgddknatasfiyngrlvdsivswskeilr-
t
qesecvcingtctvvmtdgsasgkadtkilfieegkivhtstlsgsaqhveecscyprylgvrcvcrdnwkgsn-
r
pivdinikdysivssyvcsglvgdtprkndssssshcldpnneegghgvkgwafddgndvwmgrtiseklrsgy-
e
tfkviegwsnpnsklqinrqvivdrgnrsgysgifsvegkscinrcfyvelirgrkeetevlwtsnsivvfcgt-
s gtygtgswpdgadinlmpi SEQUENCE: 44 (PA, 105p30)
medfvrqcfnpmivelaekamkeygedpkietnkfaaicthlevcfmysdfhfidergesiivesgdpnallkh-
r
feiiegrdrimawtvinsicnttgvekpkflpdlydykenrfieigvtrrevhiyylekankiksekthihifs-
f
tgeematkadytldeesrariktrlftirqemaskslwdsfrqsergeetieekfeitgtmrkladqslppnfp-
s
lenfrayvdgfepngciegklsqmskevnakiepflrttprplrlpdgplchqrskfllmdalklsiedpsheg-
e
giplydaikcmktffgwkepnivkphekginpnylmawkqvlaelqdieneekiprtknmkrtsqlkwalgenm-
a
pekvdfddckdvgdlkqydsdepeprslaswvqnefnkaceltdsswieldeigedvapiehiasmrrnyftae-
v
shcrateyimkgvyintallnascaamddfqlipmiskcrtkegrrktnlygfiikgrshlrndtdvvnfvsme-
f
sltdprlephkwekycvleigdmllrtaigqvsrpmflyvrtngtskikmkwgmemrrcllqslqqiesmieae-
s
svkekdmtkeffenksetwpigesprgveegsigkvcrtllaksvfnslyaspqlegfsaesrklllivqalrd-
n lepgtfdlgglyeaieeclindpwvllnaswfnsflthalk SEQUENCE: 45 (M1,
105p30)
mslltevetyvlsivpsgplkaeiaqrlenvfagkntdlealmewlktrpilspltkgilgfvftltvpsergl-
q
rrrfvqnalngngdpnnmdkavklyrklkreitfhgakeialsysagalascmgliynrmgavttesafglica-
t
ceqiadsqhkshrqmvtttnplirhenrmvlasttakameqmagsseqaaeamevasqarqmvqamraigthps-
s stglkndllenlqayqkrmgvqmqrfk SEQUENCE: 46 (A/Texas/1/77 PB1)
mdvnptllflkipaqnaisttfpytgdppyshgtgtgytmdtvnrthqysekgkwttntetgapqlnpidgplp-
e
dnepsgyaqtdcvleamafleeshpgifenscletmevvqqtrvdrltqgrqtydwtlnrnqpaatalantiev-
f
rsngltanesgrlidflkdvmesmdkeeieitthfqrkrrvrdnmtkkmvtqrtigkkkqrvnkrsyliraltl-
n
tmtkdaergklkrraiatpgmqirgfvyfvetlarsicekleqsglpvggnekkaklanvvrkmmtnsqdtels-
f
titgdntkwnenqnprmflamityitknqpewfrnilsiapimfsnkmarlgkgymfeskrmklrtqipaemla-
s
idlkyfnestrkkiekirpllidgtaslspgmmmgmfnmlstvlgvsilnlgqkkytkttywwdglqssddfal-
i
vnapnhegiqagvdrfyrtcklvginmskkksyinrtgtfeftsffyrygfvanfsmelpsfgvsginesadms-
i
gvtviknnminndlgpataqmalqlfikdyrytyrchrgdtqiqtrrsfelkklweqtrskagllvsdggpnly-
n
irnlhipevclkwelmdedyqgrlcnplnpfvshkeiesvnnavvmpahgpaksmeydavatthswipkrnrsi-
l
ntsqrgiledeqmyqkccnlfekffpsssyrrpvgissmveamvsraridaridfesgrikkeefseimkicst-
i eelrrqkq SEQUENCE: 47 (A/Puerto Rico/8/34 PA)
medfvrqcfnpmivelaektmkeygedlkietnkfaaicthlevcfmysdfhfineqgesiivelgdpnallkh-
r
feiiegrdrtmawtvvnsicnttgaekpkflpdlydykenrfieigvtrrevhiyylekankiksekthihifs-
f
tgeematkadytldeesrariktrlftirqemasrglwdsfrqsergeetieerfeitgtmrkladqslppnfs-
s
lenfrayvdgfepngyiegklsqmskevnariepflkttprplrlpngppcsqrskfllmdalklsiedpsheg-
e
giplydaikcmrtffgwkepnvvkphekginpnyllswkqvlaelqdieneekipktknmkktsqlkwalgenm-
a
pekvdfddckdvgdlkqydsdepelrslaswiqnefnkaceltdsswieldeigedvapiehiasmrrnyftse-
v
shcrateyimkgvyintallnascaamddfqlipmiskcrtkegrrktnlygfiikgrshlrndtdvvnfvsme-
f
sltdprlephkwekycvleigdmlirsaigqvsrpmflyvrtngtskikmkwgmemrrcllqslqqiesmieae-
s
svkekdmtkeffenksetwpigespkgveessigkvcrtllaksvfnslyaspqlegfsaesrklllivqalrd-
n lepgtfdlgglyeaieeclindpwvllnaswfnsflthals SEQUENCE: 48 (A/Puerto
Rico/8/34 NP)
masqgtkrsyeqmetdgerqnateirasvgkmiggigrfyiqmctelklsdyegrliqnsltiermvlsafder-
r
nkyleehpsagkdpkktggpiyrrvngkwmrelilydkeeirriwrqanngddataglthmmiwhsnlndatyq-
r
tralvrtgmdprmcslmqgstlprrsgaagaavkgvgtmvmelvrmikrgindrnfwrgengrktriayermcn-
i
lkgkfqtaaqkammdqvresrdpgnaefedltflarsalilrgsvahksclpacvygpavasgydferegyslv-
g idpfrllqnsqvyslirpnenpahks SEQUENCE: 49 (A/Puerto Rico/8/34 M)
mslltevetyvlsiipsgplkaeiaqrledvfagkntdlevlmewlktrpilspltkgilgfvftltvpsergl-
q
rrrfvqnalngngdpnnmdkavklyrklkreitfhgakeislsysagalascmgliynrmgavttevafglvca-
t
ceqiadsqhrshrqmvtttnplirhenrmvlasttakameqmagsseqaaeamevasqarqmvqamrtigthps-
s saglkndllenlqayqkrmgvqmqrfk SEQUENCE: 50 (HA, A/California/04/09)
mkailvvllytfatanadtlcigyhannstdtvdtvleknvtvthsvnlledkhngklcklrgvaplhlgkcni-
agw
ilgnpeceslstasswsyivetpssdngtcypgdfidyeelreqlssyssferfeifpktsswpnhdsnkgvta-
acp
hagaksfyknliwlvkkgnsypklsksyindkgkevlvlwgihhpstsadqqslyqnadtyvfvgssryskkfk-
pei
airpkvrdqegrmnyywtlvepgdkitfeatgnlvvpryafamernagsgiiisdtpvhdcnttcqtpkgaint-
slp
fqnihpitigkcpkyvkstklrlatglrnipsiqsrglfgaiagfieggwtgmvdgwygyhhqneqgsgyaadl-
kst
qnaideitnkvnsviekmntqftavgkefnhlekrienlnkkvddgfldiwtynaellvllenertldyhdsnv-
knl
yekvrsqlknnakeigngcfefyhkcdntcmesvkngtydypkyseeaklnreeidgvklestriyqilaiyst-
vas slvlvvslgaisfwmcsngslqcrici
REFERENCES
[0218] [1] WO2007/002008 [0219] [2] WO2007/124327 [0220] [3]
WO2010/070098 [0221] [4] Needleman & Wunsch (1970) J. Mol.
Biol. 48, 443-453. [0222] [5] Rice et al. (2000) Trends Genet.
16:276-277. [0223] [6] Herlocher et al. (2004) J Infect Dis
190(9):1627-30. [0224] [7] Le et al. (2005) Nature 437(7062):1108.
[0225] [8] U.S. Pat. No. 6,468,544. [0226] [9] Neumann et al.
(2005) Proc Natl Acad Sci USA 102: 16825-9 [0227] [10]
WO2009/000891 [0228] [11] U.S. provisional application No.
61/273,151 [0229] [12] Sambrook et al, Molecular Cloning: A
Laboratory Manual, 2 ed., 1989, Cold Spring Harbor Press, Cold
Spring Harbor, N.Y. [0230] [13] WO2011/012999 [0231] [14] Kistner
et al. (1998) Vaccine 16:960-8. [0232] [15] Bruhl et al. (2000)
Vaccine 19:1149-58. [0233] [16] Pau et al. (2001) Vaccine
19:2716-21. [0234] [17] http://www.atcc.org/ [0235] [18]
http://locus.umdnj.edu/ [0236] [19] WO97/37000. [0237] [20]
Halperin et al. (2002) Vaccine 20:1240-7. [0238] [21] EP-A-1260581
(WO01/64846) [0239] [22] WO2006/071563 [0240] [23] WO2005/113758
[0241] [24] WO97/37001 [0242] [25] WO02/28422. [0243] [26]
WO2005/113756. [0244] [27] Huckriede et al. (2003) Methods Enzymol
373:74-91. [0245] [28] Vaccines. (eds. Plotkins & Orenstein).
4th edition, 2004, ISBN: 0-7216-9688-0 [0246] [29] Treanor et al.
(1996) J Infect Dis 173:1467-70. [0247] [30] Keitel et al. (1996)
Clin Diagn Lab Immunol 3:507-10. [0248] [31] Herlocher et al.
(2004) J Infect Dis 190(9):1627-30. [0249] [32] Le et al. (2005)
Nature 437(7062):1108. [0250] [33] WO2008/068631. [0251] [34]
Gennaro (2000) Remington: The Science and Practice of Pharmacy.
20th edition, ISBN: 0683306472. [0252] [35] Banzhoff (2000)
Immunology Letters 71:91-96. [0253] [36] Nony et al. (2001) Vaccine
27:3645-51. [0254] [37] EP-B-0870508. [0255] [38] U.S. Pat. No.
5,948,410. [0256] [39] WO2007/052163. [0257] [40] WO2007/052061
[0258] [41] WO90/14837. [0259] [42] Podda (2001) Vaccine 19:
2673-2680. [0260] [43] Vaccine Design: The Subunit and Adjuvant
Approach (eds. Powell & Newman) Plenum Press 1995 (ISBN
0-306-44867-X). [0261] [44] Vaccine Adjuvants: Preparation Methods
and Research Protocols (Volume 42 of Methods in Molecular Medicine
series). ISBN: 1-59259-083-7. Ed. O'Hagan. [0262] [45]
WO2008/043774. [0263] [46] Allison & Byars (1992) Res Immunol
143:519-25. [0264] [47] Hariharan et al. (1995) Cancer Res
55:3486-9. [0265] [48] US-2007/014805. [0266] [49] US-2007/0191314.
[0267] [50] Suli et al. (2004) Vaccine 22(25-26):3464-9. [0268]
[51] WO95/11700. [0269] [52] U.S. Pat. No. 6,080,725. [0270] [53]
WO2005/097181. [0271] [54] WO2006/113373. [0272] [55] Potter &
Oxford (1979) Br Med Bull 35: 69-75. [0273] [56] Greenbaum et al.
(2004) Vaccine 22:2566-77. [0274] [57] Piascik (2003) J Am Pharm
Assoc (Wash DC). 43:728-30. [0275] [58] Mann et al. (2004) Vaccine
22:2425-9. [0276] [59] Halperin et al. (1979) Am J Public Health
69:1247-50. [0277] [60] Herbert et al. (1979) J Infect Dis
140:234-8. [0278] [61] Chen et al. (2003) Vaccine 21:2830-6. [0279]
[62] Current Protocols in Molecular Biology (F. M. Ausubel et al.,
eds., 1987) Supplement 30. [0280] [63] Smith & Waterman (1981)
Adv. Appl. Math. 2: 482-489.
Sequence CWU 1
1
5012201DNAInfluenza A virus 1gattcgaaat ggaagatttt gtgcgacaat
gcttcaatcc gatgattgtc gagcttgcgg 60aaaaggcaat gaaagagtat ggagaggacc
tgaaaatcga aacaaacaaa tttgcagcaa 120tatgcactca cttggaagta
tgcttcatgt attcagattt tcatttcatc aatgagcaag 180gcgaatcaat
aatagtagag cctgaggacc caaatgcact tttaaagcac agatttgaga
240taatagaggg acgagatcgt acaatggcat ggacagttgt aaacagtatt
tgcaacacca 300caggagctga gaaaccaaag tttctgccag atctgtatga
ttacaaagag aatagattca 360tcgagattgg agtgacaagg agggaagttc
acatatacta tctggaaaag gccaacaaaa 420ttaaatctga gaagacacac
attcacattt tctcattcac tggcgaagaa atggccacaa 480aggccgatta
cactctcgat gaagaaagca gggctaggat taaaaccaga ctattcacca
540taagacaaga aatggcaagc agaggtcttt gggactcctt tcgtcagtcc
gaaagaggcg 600aagaaacaat tgaagaaaga tttgaaatca cagggacaat
gcgcaggctc gctgaccaaa 660gccttccgcc gaacttctcc tgcattgaga
attttagagc ctatgtggat ggatttgaac 720cgaacggcta cattgagggc
aagctttctc aaatgtccaa agaagtaaat gctagaattg 780agcctttttt
gaaaacaaca ccacgaccaa ttagacttcc ggatgggcct ccttgttttc
840agcggtcaaa attcctgctg atggattctt taaaattaag cattgaggat
ccaaatcatg 900aaggagaggg aataccacta tatgatgcaa tcaagtgtat
gagaacattc tttggatgga 960aagaaccctc tgttgtcaag ccacacggga
agggaataaa tccgaattat ctgctgtcat 1020ggaagcaggt attggaagag
ctgcaggaca ttgagagtga ggagaagatt ccaagaacaa 1080aaaacatgaa
aaaaacgagt cagctaaagt gggcacttgg tgagaacatg gcaccagaga
1140aggtggattt tgatgactgt aaagatataa gcgatttgaa gcaatatgat
agtgacgaac 1200ctgaattaag gtcattttca agttggatcc agaatgagtt
caacaaggca tgcgagctga 1260ccgattcaat ctggatagag ctcgatgaga
ttggagaaga tgtggccccg attgaacaca 1320ttgcaagcat gagaagaaat
tacttcacag ctgaggtgtc ccattgcaga gccacagaat 1380atataatgaa
gggggtatac attaatactg ctttgcttaa tgcatcctgt gcagcaatgg
1440atgatttcca actaattccc atgataagca aatgtagaac taaagaggga
aggagaaaga 1500ccaatttgta cggcttcatc gtaaaaggaa gatctcactt
aaggaatgac accgatgtgg 1560taaactttgt gagcatggag ttttccctca
ctgacccaag acttgagcca cacaaatggg 1620agaagtactg tgttcttgag
ataggagata tgcttctaag gagtgcaata ggccaagtgt 1680caaggcccat
gttcttgtat gtaaggacaa atggaacctc aaaaattaaa atgaaatggg
1740gaatggagat gaggcgttgc ctcctccaat cccttcaaca aatagagagc
atgattgaag 1800ctgagtcctc cgtcaaggag aaagacatga caaaagagtt
ttttgagaat agatcagaaa 1860catggcccat tggagagtca ccaaaaggag
tggaagaagg ttccattggg aaagtatgca 1920ggacactatt ggctaagtca
gtattcaata gtctgtatgc atctccacaa ttagaaggat 1980tttcagctga
gtcaagaaag ttgctcctca ttgttcaggc tcttagggac aatctggaac
2040ctgggacctt tgatcttggg gggctatatg aagcaattga ggagtgcctg
attaatgatc 2100cctgggtttt gcttaatgct tcttggttca actccttcct
aacacatgca ttgagatagc 2160tggggcaatg ctactattta ctatccatac
tgtccaaaaa a 220122301DNAInfluenza A virus 2aatggatgtc aatccgacat
tacttttctt aaaagtgcca gcacaaaatg ctataagcac 60aacttttcct tatactggtg
accctcctta cagccatggg acaggaacag ggtacaccat 120ggatacagtc
aacaggacac atcagtactc agaaagagga agatggacaa aaaataccga
180aactggagca ccgcaactca acccaattga tgggccacta ccaaaagaca
atgaaccaag 240tggctatgcc caaacagatt gtgtattaga agcaatggct
ttccttgagg aatcccatcc 300tggtattttt gaaaactctt gtattgaaac
aatggaggtt gttcagcaaa caagggtgga 360caaactgaca caaggcagac
agacctatga ctggactcta aataggaacc agcctgctgc 420cacagcattg
gccaacacta tagaagtgtt cagatcaaac ggcctcatag caaatgaatc
480tgggaggcta atagacttcc ttaaagatgt aatggagtcg atggacagag
acgaagtaga 540gatcacaact cattttcaaa gaaagaggag agtgagagac
aatgtaacta aaaaaatggt 600gacccaaaga acaataggca aaaagaaaca
taaattagac aaaagaagtt acctaattag 660ggcattaacc ctgaacacaa
tgaccaaaga tgctgagagg gggaaactaa aacgcagagc 720aattgcaacc
ccaggaatgc aaataagggg gtttgtatac tttgttgaga cactggcaag
780aagcatatgt gaaaagcttg aacaatcagg gttgccagtt ggaggaaatg
aaaagaaagc 840aaagttagca aatgttgtaa ggaagatgat gaccaactcc
caggacactg aaatttcttt 900caccatcact ggagataaca caaaatggaa
cgaaaatcaa aaccctagaa tgttcttggc 960catgatcaca tatataacca
aaaatcagcc tgaatggttc agaaatattc taagtattgc 1020tccaataatg
ttttcaaaca aaatggcgag actaggtaag gggtacatgt ttgaaagcaa
1080gagtatgaaa ctgagaactc aaatacctgc agagatgcta gccaacatag
atttgaaata 1140tttcaatgat tcaactaaaa agaaaattga aaaaatccgg
ccattattaa tagatggaac 1200tgcatcattg agtcctggaa tgatgatggg
catgttcaat atgttaagca ccgtcttggg 1260cgtctccatt ctgaatcttg
ggcaaaagag atacaccaag actacttact ggtgggatgg 1320tcttcaatcg
tctgatgatt ttgctctgat tgtgaatgca cccaactatg caggaattca
1380agctggagtt gacaggtttt atcgaacctg taagctgctc ggaattaata
tgagcaaaaa 1440gaagtcttac ataaacagaa caggtacctt tgagttcacg
agctttttct atcgttatgg 1500gtttgttgcc aatttcagca tggagcttcc
tagttttggg gtgtctgggg tcaatgaatc 1560tgcagacatg agtattggag
tcactgtcat caaaaacaat atgataaaca atgaccttgg 1620cccagcaact
gctcaaatgg cccttcagtt atttataaaa gattacaggt acacgtatcg
1680atgccacaga ggtgacacac aaatacaaac ccggagatca tttgagataa
agaaactatg 1740ggaccaaacc cgctccaaag ctgggctgtt ggtctctgat
ggaggcccca atttatataa 1800cattagaaat ctccatattc ctgaagtctg
cttgaaatgg gagttgatgg atgaggatta 1860ccaggggcgt ttatgcaacc
cattgaaccc gtttgtcagt cataaagaga ttgaatcagt 1920gaacaatgca
gtgatgatgc cggcacatgg tccagccaaa aatatggagt atgacgctgt
1980tgcaacaaca cactcctggg ttcccaaaag gaatcgatcc attttgaata
cgagccaaag 2040ggggatactt gaggatgagc aaatgtatca gaggtgctgc
aatttatttg aaaaattctt 2100cccaagtagc tcatacagaa gaccagttgg
aatatccagt atggtagagg ctatggtttc 2160cagagcccga attgatgcac
ggattgattt cgaatctgga aggataaaaa aagaggaatt 2220cgctgagatc
atgaagacct gttccaccat tgaagacctc agacggcaaa aatagggaat
2280ttggcttgtc cttcatgaaa a 230132299DNAInfluenza A virus
3aatatggaaa gaataaaaga gctaaggaat ctgatgtcac aatctcgcac tcgcgagata
60cttacaaaaa ctactgtaga ccacatggcc ataatcaaga aatacacatc aggaagacag
120gagaaaaacc catcacttag aatgaaatgg atgatggcaa tgaaataccc
aattacagca 180gataaaagga taacggaaat gattcctgaa agaaatgagc
aaggacagac attatggagt 240aaagtgaatg atgccggatc agaccgagtg
atgatatcac ccctggctgt gacatggtgg 300aacagaaatg gaccagtggc
aagtactatt cactatccaa aaatctacaa aacttacttt 360gaaaaggttg
aaaggttaaa acatggaacc tttggccctg tacactttag aaaccaagtc
420aaaatacgcc gaagagtcga cataaatcct ggtcatgcag acctcagcgc
caaggaggca 480caggatgtaa ttatggaagt tgttttccct aatgaagtgg
gagccagaat actaacatca 540gaatcgcaat taacgataac caaggagaaa
aaagaagaac tccagaattg caaaatttcc 600cctttgatgg ttgcatacat
gttagagagg gaacttgtcc gcaaaacgag atttctcccg 660gttgctggtg
gaacaagcag tgtgtacatt gaagttttgc atttaacaca ggggacatgc
720tgggagcaga tgtacactcc aggtggggag gtgaggaatg atgatgttga
tcaaagccta 780attattgctg ctaggaacat agtgagaaga gctgcagtat
cagcagatcc actagcatct 840ttattagaaa tgtgccatag cacacagatt
ggtgggacaa ggatggtgga tattctcagg 900caaaatccaa cagaagaaca
agctgtggat atatgcaaag cagcaatggg gctgagaatc 960agttcatcct
tcagttttgg cggattcaca tttaagagaa caagtggatc atcagtcaaa
1020agggaggaag aagtgctcac gggcaatctg caaacattga agctaactgt
gcatgaggga 1080tatgaagagt tcacaatggt tgggaaaagg gcaacagcta
tactcagaaa agcaaccagg 1140agattgattc aactaatagt gagtggaaga
gacgaacagt caatagtcga agcaatagtt 1200gtagcaatgg tattctcaca
agaagattgc atggtaaaag cagttagagg tgatctgaat 1260ttcgttaata
gagcgaatca gcggttgaat cccatgcatc aacttttgag acattttcag
1320aaggatgcta aagtactttt cttaaattgg ggaattgaac ctatcgacaa
tgtgatggga 1380atgattggga tattacctga tatgactcca agtaccgaga
tgtcaatgag aggagtgaga 1440gtcagcaaaa tgggtgtaga tgaatactcc
aatgctgaaa gggtagtggt gagcattgac 1500cgttttttga gagtccggga
ccaaagagga aatgtactac tgtctccaga ggaagtcagt 1560gaaacacagg
gaacagagaa actgacaata acttactctt catcaatgat gtgggagatt
1620aatggccctg agtcagtgtt gatcaatacc tatcagtgga tcatcagaaa
ctgggagact 1680gttaaaattc agtggtctca gaaccctaca atgctataca
ataaaatgga attcgagcca 1740tttcagtctc tagtccctaa ggccattaga
ggccaataca gtgggtttgt tagaactcta 1800tttcaacaaa tgagggatgt
gcttgggacc tttgacacaa ctcagataat aaaacttctt 1860ccctttgcag
ccgctccacc aaagcaaagt agaatgcaat tctcatcatt gactgtgaat
1920gtgaggggat caggaatgag aatacttgta aggggtaatt ctccagtatt
caactacaac 1980aagaccacta agagactcac agtcctcgga aaggatgctg
gcactttaac tgaagaccca 2040gatgaaggca cagctggagt ggaatctgct
gttctaaggg gattcctcat tctaggcaaa 2100gaagatagaa gatatgggcc
agcattaagc atcaatgaat tgagcaacct tgcgaaaggg 2160gaaaaagcta
atgtgctaat tgggcaaggg gacgtagtgt tggtaatgaa acgaaaacgg
2220gactctagca tacttactga cagccagaca gcgaccaaaa gaattcggat
ggccatcaat 2280taatttcgaa taatttaaa 229941527DNAInfluenza A virus
4atcactcact gagtgacatc aaagtcatgg cgtcccaagg caccaaacgg tcttacgaac
60agatggagac tgatggggaa cgccagaatg caactgaaat cagagcatcc gtcggaagaa
120tgattggtgg aattgggcga ttctacatcc aaatgtgcac cgagcttaaa
ctcaatgatt 180atgagggacg actgatccag aacagcttga caatagagag
aatggtgctc tctgcttttg 240atgagaggag gaataaatat ctggaagaac
atcccagcgc ggggaaagat cctaagaaaa 300ctggaggacc catatacaag
agagtagatg gaaagtgggt gagggaactc gtcctttatg 360acaaagaaga
aataaggcgg atttggcgcc aagccaacaa tggtgatgat gcaacggctg
420gtttgactca cattatgatc tggcattcta atttgaatga tacaacttac
cagaggacaa 480gagctcttgt ccgcaccgga atggatccca ggatgtgctc
tttgatgcaa ggttcaactc 540tccctagaag atctggagca gcaggcgctg
cagtcaaagg agttgggaca atggtgttgg 600agttaatcag gatgatcaaa
cgtgggatca atgaccgaaa cttctggagg ggtgagaatg 660gaagaaaaac
aaggattgct tatgagagaa tgtgcaacat tctcaaagga aaatttcaaa
720cagctgcaca aaaagcaatg atggatcaag tgagagaaag ccggaaccca
ggaaatgctg 780agatcgaaga tctcactttt ctggcacggt ctgcactcat
attaagaggg tcagttgctc 840acaagtcttg cctgcctgcc tgtgtgtatg
gaccagccgt agccagtggg tacgacttcg 900aaaaagaggg atactctttg
gtaggggtag acccttttaa actgcttcaa accagtcagg 960tatacagcct
aatcagacca aacgagaatc ccgcacacaa gagtcagttg gtgtggatgg
1020catgcaattc tgctgcattt gaagatctaa gagtgtcaag cttcatcaga
gggacaagag 1080tacttccaag ggggaagctc tccactagag gagtacaaat
tgcttcaaat gaaaacatgg 1140atgctattgt atcaagtact cttgaactga
gaagcagata ctgggccata agaaccagaa 1200gtggagggaa cactaatcaa
caaagggcct ctgcgggcca aatcagcaca caacctacgt 1260tttctgtgca
gagaaacctc ccatttgaca aaacaaccat catggcagca ttcactggga
1320atacggaggg aagaacatca gacatgaggg cagaaatcat aaagatgatg
gaaagtgcaa 1380gaccagaaga agtgtccttc caggggcggg gagtctttga
gctctcggac gaaagggcaa 1440cgaacccgat cgtgccctcc tttgacatga
gtaatgaagg atcttatttc ttcggagaca 1500atgcagagga gtacgacaat taatgaa
15275984DNAInfluenza A virus 5gatgagtctt ctaaccgagg tcgaaacgta
cgttctctct atcgtcccgt caggccccct 60caaagccgag atcgcacaga gacttgaaaa
tgtctttgct ggaaagaata ccgatcttga 120ggctctcatg gaatggctaa
agacaagacc aatcctgtca cctctgacta aggggatttt 180aggatttgtg
ttcacgctca ccgtgcccag tgagcgagga ctgcagcgta gacgctttgt
240ccaaaatgcc cttaatggga atggggatcc aaataatatg gacagagcag
ttaaactgta 300tcgaaagctt aagagggaga taacattcca tggggccaaa
gaaatagcac tcagttattc 360tgctggtgca cttgccagtt gtatgggact
catatacaac aggatggggg ctgtgaccac 420cgaatcagca tttggcctta
tatgcgcaac ctgtgaacag attgccgact cccagcataa 480gtctcatagg
caaatggtaa caacaaccaa cccattaata agacatgaga acagaatggt
540tctggccagc actacagcta aggctatgga gcaaatggct ggatcgagtg
aacaagcagc 600tgaggccatg gaggttgcta gtcaggccag gcagatggtg
caggcaatga gagccattgg 660gactcatcct agctctagca ctggtctgaa
aaatgatctc cttgaaaatt tgcaggccta 720tcagaaacga atgggggtgc
agatgcaacg attcaagtga tcctcttgtt gttgccgcaa 780gtataattgg
gattgtgcac ctgatattgt ggattattga tcgccttttt tccaaaagca
840tttatcgtat ctttaaacac ggtttaaaaa gagggccttc tacggaagga
gtaccagagt 900ctatgaggga agaatatcga gaggaacagc agaatgctgt
ggatgctgac gatggtcatt 960ttgtcagcat agagctagag taaa
9846844DNAInfluenza A virus 6atggattccc acactgtgtc aagctttcag
gtagattgct tcctttggca tgtccgcaaa 60caagttgcag accaagatct aggcgatgcc
ccattccttg atcggcttcg ccgagatcag 120aagtctctaa agggaagagg
cagcactctc ggtctgaaca tcgaaacagc cacttgtgtt 180ggaaagcaaa
tagtagagag gattctgaaa gaagaatccg atgaggcatt taaaatgacc
240atggcctccg cacttgcttc gcggtaccta actgacatga ctattgaaga
aatgtcaagg 300gactggttca tgctcatgcc caagcagaaa gtggctggcc
ctctttgtgt cagaatggac 360caggcgataa tggataagaa catcatactg
aaagcgaatt tcagtgtgat ttttgaccgg 420ttggagaatc tgacattact
aagggctttc accgaagagg gagcaattgt tggcgaaatt 480tcaccattgc
cttctcttcc aggacatact aatgaggatg tcaaaaatgc aattggggtc
540ctcatcgggg gacttgaatg gaatgataac acagttcgag tctctgaaac
tctacagaga 600ttcgcttgga gaagcagtaa tgagactggg ggacctccat
tcactccaac acagaaacgg 660aaaatggcgg gaacaattag gtcagaagtt
tgaagaaata agatggctga ttgaagaagt 720gaggcataaa ttgaagacga
cagagaatag ttttgagcaa ataacattta tgcaagcatt 780acagctattg
tttgaagtgg aacaagagat tagaacgttt tcgtttcagc ttatttaatg 840ataa
84471728DNAInfluenza A virus 7ccaaaatgaa agcaaaacta ctggtcctgt
tatgtacatt tacagctaca tatgcagaca 60caatatgtat aggctaccat gccaacaact
caaccgacac tgttgacaca gtacttgaga 120agaatgtgac agtgacacac
tctgtcaacc tacttgagga cagtcacaat ggaaaactat 180gtctactaaa
aggaatagcc ccactacaat tgggtaattg cagcgttgcc ggatggatct
240taggaaaccc agaatgcgaa ttactgattt ccaaggaatc atggtcctac
attgtagaaa 300caccaaatcc tgagaatgga acatgttacc cagggtattt
cgccgactat gaggaactga 360gggagcaatt gagttcagta tcttcatttg
agagattcga aatattcccc aaagaaagct 420catggcccaa ccacaccgta
accggagtat cagcatcatg ctcccataat gggaaaagca 480gtttttacag
aaatttgcta tggctgacgg ggaagaatgg tttgtaccca aacctgagca
540agtcctatgt aaacaacaaa gagaaagaag tccttgtact atggggtgtt
catcacccgc 600ctaacatagg gaaccaaagg gccctctatc atacagaaaa
tgcttatgtc tctgtagtgt 660cttcacatta tagcagaaga ttcaccccag
aaatagccaa aagacccaaa gtaagagatc 720aggaaggaag aatcaactac
tactggactc tgctggaacc tggggataca ataatatttg 780aggcaaatgg
aaatctaata gcgccatggt atgcttttgc actgagtaga ggctttggat
840caggaatcat cacctcaaat gcaccaatgg atgaatgtga tgcgaagtgt
caaacacctc 900agggagctat aaacagcagt cttcctttcc agaatgtaca
cccagtcaca ataggagagt 960gtccaaagta tgtcaggagt gcaaaattaa
ggatggttac aggactaagg aacatcccat 1020ccattcaatc cagaggtttg
tttggagcca ttgccggttt cattgaaggg gggtggactg 1080gaatggtaga
tgggtggtat ggttatcatc atcagaatga gcaaggatct ggctatgctg
1140cagatcaaaa aagtacacaa aatgccatta acgggattac aaacaaggtg
aattctgtaa 1200ttgagaaaat gaacactcaa ttcacagctg tgggcaaaga
attcaacaaa ttggaaagaa 1260ggatggaaaa cttaaataaa aaagttgatg
atgggtttct agacatttgg acatataatg 1320cagaattgtt ggttctactg
gaaaatgaaa ggactttgga tttccatgac tccaatgtga 1380agaatctgta
tgagaaagta aaaagccaat taaagaataa tgccaaagaa ataggaaacg
1440ggtgttttga attctatcac aagtgtaaca atgaatgcat ggagagtgtg
aaaaatggaa 1500cttatgacta tccaaaatat tccgaagaat caaagttaaa
cagggagaaa attgatggag 1560tgaaattgga atcaatggga gtctatcaga
ttctggcgat ctactcaact gtcgccagtt 1620ccctggttct tttggtctcc
ctgggggcaa tcagcttctg gatgtgttcc aatgggtctt 1680tgcagtgtag
aatatgcatc tgagaccaga atttcagaaa tataagaa 172881414DNAInfluenza A
virus 8aatgaatcca aatcaaaaaa taataaccat tggatcaatc agtatagcaa
tcggaataat 60tagtctaatg ttgcaaatag gaaatattat ttcaatatgg gctagtcact
caatccaaac 120tggaagtcaa aaccacactg gagtatgcaa ccaaagaatc
atcacatatg aaaacagcac 180ctgggtgaat cacacatatg ttaatattaa
caacactaat gttgttgctg gaaaggacaa 240aacttcagtg acattggccg
gcaattcatc tctttgttct atcagtggat gggctatata 300cacaaaagac
aacagcataa gaattggctc caaaggagat gtttttgtca taagagaacc
360tttcatatca tgttctcact tggaatgcag aacctttttt ctgacccaag
gtgctctatt 420aaatgacaaa cattcaaatg ggaccgttaa ggacagaagt
ccttataggg ccttaatgag 480ctgtcctcta ggtgaagctc cgtccccata
caattcaaag tttgaatcag ttgcatggtc 540agcaagcgca tgccatgatg
gcatgggctg gttaacaatc ggaatttctg gtccagacaa 600tggagctgtg
gctgtactaa aatacaacgg cataataact gaaaccataa aaagttggaa
660aaagcgaata ttaagaacac aagagtctga atgtgtctgt gtgaacgggt
catgtttcac 720cataatgacc gatggcccga gtaatggggc cgcctcgtac
aaaatcttca agatcgaaaa 780ggggaaggtt actaaatcaa tagagttgaa
tgcacccaat tttcattatg aggaatgttc 840ctgttaccca gacactggca
cagtgatgtg tgtatgcagg gacaactggc atggttcaaa 900tcgaccttgg
gtgtctttta atcaaaacct ggattatcaa ataggataca tctgcagtgg
960ggtgttcggt gacaatccgc gtcccaaaga tggagagggc agctgtaatc
cagtgactgt 1020tgatggagca gacggagtaa aggggttttc atacaaatat
ggtaatggtg tttggatagg 1080aaggactaaa agtaacagac ttagaaaggg
gtttgagatg atttgggatc ctaatggatg 1140gacagatacc gacagtgatt
tctcagtgaa acaggatgtt gtggcaataa ctgattggtc 1200agggtacagc
ggaagtttcg ttcaacatcc tgagttaaca ggattggact gtataagacc
1260ttgcttctgg gttgagttag tcagaggact gcctagagaa aatacaacaa
tctggactag 1320tgggagcagc atttcttttt gtggcgtaaa tagtgatact
gcaaactggt cttggccaga 1380cggtgctgag ttgccgttca ccattgacaa gtag
141492233DNAInfluenza A virus 9agcgaaagca ggtactgatc caaaatggaa
gattttgtgc gacaatgctt caatccgatg 60attgtcgagc ttgcggaaaa aacaatgaaa
gagtatgggg aggacctgaa aatcgaaaca 120aacaaatttg cagcaatatg
cactcacttg gaagtatgct tcatgtattc agattttcac 180ttcatcaatg
agcaaggcga gtcaataatc gtagaacttg gtgatccaaa tgcacttttg
240aagcacagat ttgaaataat cgagggaaga gatcgcacaa tggcctggac
agtagtaaac 300agtatttgca acactacagg ggctgagaaa ccaaagtttc
taccagattt gtatgattac 360aaggagaata gatttatcga aattggagta
acaaggagag aagttcacat atactatctg 420gaaaaggcca ataaaattaa
atctgagaaa acacacatcc acattttctc gttcactggg 480gaagaaatgg
ccacaaaggc agactacact ctcgatgaag aaagcagggc taggatcaaa
540accagactat tcaccataag acaagaaatg gccagcagag gcctctggga
ttcctttcgt 600cagtccgaga gaggagaaga gacaattgaa gaaaggtttg
aaatcacagg aacaatgcgc 660aagcttgccg accaaagtct cccgccgaac
ttctccagcc ttgaaaattt tagagcctat 720gtggatggat tcgaaccgaa
cggctacatt gagggcaagc tgtctcaaat gtccaaagaa 780gtaaatgcta
gaattgaacc ttttttgaaa acaacaccac gaccacttag acttccgaat
840gggcctccct gttctcagcg gtccaaattc ctgctgatgg atgccttaaa
attaagcatt 900gaggacccaa gtcatgaagg agagggaata ccgctatatg
atgcaatcaa atgcatgaga 960acattctttg gatggaagga acccaatgtt
gttaaaccac acgaaaaggg aataaatcca 1020aattatcttc tgtcatggaa
gcaagtactg gcagaactgc aggacattga gaatgaggag 1080aaaattccaa
agactaaaaa tatgaagaaa acaagtcagc taaagtgggc acttggtgag
1140aacatggcac cagaaaaggt agactttgac gactgtaaag atgtaggtga
tttgaagcaa 1200tatgatagtg atgaaccaga attgaggtcg cttgcaagtt
ggattcagaa tgagtttaac
1260aaggcatgcg aactgacaga ttcaagctgg atagagctcg atgagattgg
agaagatgtg 1320gctccaattg aacacattgc aagcatgaga aggaattatt
tcacatcaga ggtgtctcac 1380tgcagagcca cagaatacat aatgaagggg
gtgtacatca atactgcctt gcttaatgca 1440tcttgtgcag caatggatga
tttccaatta attccaatga taagcaagtg tagaactaag 1500gagggaaggc
gaaagaccaa cttgtatggt ttcatcataa aaggaagatc ccacttaagg
1560aatgacaccg acgtggtaaa ctttgtgagc atggagtttt ctctcactga
cccaagactt 1620gaaccacata aatgggagaa gtactgtgtt cttgagatag
gagatatgct tataagaagt 1680gccataggcc aggtttcaag gcccatgttc
ttgtatgtga gaacaaatgg aacctcaaaa 1740attaaaatga aatggggaat
ggagatgagg cgttgcctcc tccagtcact tcaacaaatt 1800gagagtatga
ttgaagctga gtcctctgtc aaagagaaag acatgaccaa agagttcttt
1860gagaacaaat cagaaacatg gcccattgga gagtccccca aaggagtgga
ggaaagttcc 1920attgggaagg tctgcaggac tttattagca aagtcggtat
tcaacagctt gtatgcatct 1980ccacaactag aaggattttc agctgaatca
agaaaactgc ttcttatcgt tcaggctctt 2040agggacaacc ttgaacctgg
gacctttgat cttggggggc tatatgaagc aattgaggag 2100tgcctgatta
atgatccctg ggttttgctt aatgcttctt ggttcaactc cttccttaca
2160catgcattga gttagttgtg gcagtgctac tatttgctat ccatactgtc
caaaaaagta 2220ccttgtttct act 2233102341DNAInfluenza A virus
10agcgaaagca ggcaaaccat ttgaatggat gtcaatccga ccttactttt cttaaaagtg
60ccaacacaaa atgctataag cacaactttc ccttatactg gagaccctcc ttacagccat
120gggacaggaa caggatacac catggatact gtcaacagga cacatcagta
ctcagaaaag 180ggaagatgga caacaaacac cgaaactgga gcaccgcaac
tcaacccgat tgatgggcca 240ctgccagaag acaatgaacc aagtggttat
gcccaaacag attgtgtatt ggaggcgatg 300gctttccttg aggaatccca
tcctggtatt tttgaaaact cgtgtattga aacgatggag 360gttgttcagc
aaacacgagt agacaagctg acacaaggcc gacagaccta tgactggact
420ctaaatagaa accaacctgc tgcaacagca ttggccaaca caatagaagt
gttcagatca 480aatggcctca cggccaatga gtctggaagg ctcatagact
tccttaagga tgtaatggag 540tcaatgaaca aagaagaaat ggggatcaca
actcattttc agagaaagag acgggtgaga 600gacaatatga ctaagaaaat
gataacacag agaacaatgg gtaaaaagaa gcagagattg 660aacaaaagga
gttatctaat tagagcattg accctgaaca caatgaccaa agatgctgag
720agagggaagc taaaacggag agcaattgca accccaggga tgcaaataag
ggggtttgta 780tactttgttg agacactggc aaggagtata tgtgagaaac
ttgaacaatc agggttgcca 840gttggaggca atgagaagaa agcaaagttg
gcaaatgttg taaggaagat gatgaccaat 900tctcaggaca ccgaactttc
tttcaccatc actggagata acaccaaatg gaacgaaaat 960cagaatcctc
ggatgttttt ggccatgatc acatatatga ccagaaatca gcccgaatgg
1020ttcagaaatg ttctaagtat tgctccaata atgttctcaa acaaaatggc
gagactggga 1080aaagggtata tgtttgagag caagagtatg aaacttagaa
ctcaaatacc tgcagaaatg 1140ctagcaagca tcgatttgaa atatttcaat
gattcaacaa gaaagaagat tgaaaaaatc 1200cgaccgctct taatagaggg
gactgcatca ttgagccctg gaatgatgat gggcatgttc 1260aatatgttaa
gcactgtatt aggcgtctcc atcctgaatc ttggacaaaa gagatacacc
1320aagactactt actggtggga tggtcttcaa tcctctgacg attttgctct
gattgtgaat 1380gcacccaatc atgaagggat tcaagccgga gtcgacaggt
tttatcgaac ctgtaagcta 1440cttggaatca atatgagcaa gaaaaagtct
tacataaaca gaacaggtac atttgaattc 1500acaagttttt tctatcgtta
tgggtttgtt gccaatttca gcatggagct tcccagtttt 1560ggggtgtctg
ggatcaacga gtcagcggac atgagtattg gagttactgt catcaaaaac
1620aatatgataa acaatgatct tggtccagca acagctcaaa tggcccttca
gttgttcatc 1680aaagattaca ggtacacgta ccgatgccat agaggtgaca
cacaaataca aacccgaaga 1740tcatttgaaa taaagaaact gtgggagcaa
acccgttcca aagctggact gctggtctcc 1800gacggaggcc caaatttata
caacattaga aatctccaca ttcctgaagt ctgcctaaaa 1860tgggaattga
tggatgagga ttaccagggg cgtttatgca acccactgaa cccatttgtc
1920agccataaag aaattgaatc aatgaacaat gcagtgatga tgccagcaca
tggtccagcc 1980aaaaacatgg agtatgatgc tgttgcaaca acacactcct
ggatccccaa aagaaatcga 2040tccatcttga atacaagtca aagaggagta
cttgaggatg aacaaatgta ccaaaggtgc 2100tgcaatttat ttgaaaaatt
cttccccagc agttcataca gaagaccagt cgggatatcc 2160agtatggtgg
aggctatggt ttccagagcc cgaattgatg cacggattga tttcgaatct
2220ggaaggataa agaaagaaga gttcactgag atcatgaaga tctgttccac
cattgaagag 2280ctcagacggc aaaaatagtg aatttagctt gtccttcatg
aaaaaatgcc ttgtttctac 2340t 2341112341DNAInfluenza A virus
11agcgaaagca ggtcaattat attcaatatg gaaagaataa aagaactaag aaatctaatg
60tcgcagtctc gcacccgcga gatactcaca aaaaccaccg tggaccatat ggccataatc
120aagaagtaca catcaggaag acaggagaag aacccagcac ttaggatgaa
atggatgatg 180gcaatgaaat atccaattac agcagacaag aggataacgg
aaatgattcc tgagagaaat 240gagcaaggac aaactttatg gagtaaaatg
aatgatgccg gatcagaccg agtgatggta 300tcacctctgg ctgtgacatg
gtggaatagg aatggaccaa taacaaatac agttcattat 360ccaaaaatct
acaaaactta ttttgaaaga gtagaaaggc taaagcatgg aacctttggc
420cctgtccatt ttagaaacca agtcaaaata cgtcggagag ttgacataaa
tcctggtcat 480gcagatctca gtgccaagga ggcacaggat gtaatcatgg
aagttgtttt ccctaacgaa 540gtgggagcca ggatactaac atcggaatcg
caactaacga taaccaaaga gaagaaagaa 600gaactccagg attgcaaaat
ttctcctttg atggttgcat acatgttgga gagagaactg 660gtccgcaaaa
cgagattcct cccagtggct ggtggaacaa gcagtgtgta cattgaagtg
720ttgcatttga ctcaaggaac atgctgggaa cagatgtata ctccaggagg
ggaagtgagg 780aatgatgatg ttgatcaaag cttgattatt gctgctagga
acatagtgag aagagctgca 840gtatcagcag atccactagc atctttattg
gagatgtgcc acagcacaca gattggtgga 900attaggatgg tagacatcct
taggcagaac ccaacagaag agcaagccgt ggatatatgc 960aaggctgcaa
tgggactgag aattagctca tccttcagtt ttggtggatt cacatttaag
1020agaacaagcg gatcatcagt caagagagag gaagaggtgc ttacgggaaa
tcttcaaaca 1080ttgaagataa gagtgcatga gggatatgaa gagttcacaa
tggttgggag aagagcaaca 1140gccatactca gaaaagcaac caggagattg
attcagctga tagtgagtgg gagagacgaa 1200cagtcgattg ccgaagcaat
aattgtggcc atggtatttt cacaagagga ttgtatgata 1260aaagcagtca
gaggtgatct gaatttcgtc aatagggcga atcagcgatt gaatcctatg
1320catcaacttt taagacattt tcagaaggat gcgagagtgc tttttcaaaa
ttggggagtt 1380gaacctatcg acaatgtgat gggaatgatt gggatattgc
ccgacatgac tccaagcatc 1440gagatgtcaa tgagaggagt gagaatcagc
aaaatgggtg tagatgagta ctccagcacg 1500gagagggtag tggtgagcat
tgaccgtttt ttgagaatcc gggaccaacg aggaaatgta 1560ctactgtctc
ccgaggaggt cagtgaaaca cagggaacag agaaactgac aataacttac
1620tcatcgtcaa tgatgtggga gattaatggt cctgaatcag tattggtcaa
tacctatcaa 1680tggatcatca gaaactggga aactgttaaa attcagtggt
cccagaaccc tacaatgcta 1740tacaataaaa tggaatttga accatttcag
tctttagtac ctaaggccat tagaggccaa 1800tacagtgggt ttgtaagaac
tctgttccaa caaatgaggg atgtgcttgg gacatttgat 1860accgcacaga
taataaaact tcttcccttc gcagccgctc caccaaagca aagtagaatg
1920cagttctcct catttactgt gaatgtgagg ggatcaggaa tgagaatact
tgtaaggggc 1980aattctcctg tattcaacta taacaaggcc acgaagagac
tcacagttct cggaaaggat 2040gctggcactt taactgaaga cccagatgaa
ggcacagctg gagtggagtc cgctgttctg 2100aggggattcc tcattctggg
caaagaagac aagagatatg ggccagcact aagcatcaat 2160gaactgagca
accttgcgaa aggagagaag gctaatgtgc taattgggca aggagacgtg
2220gtgttggtaa tgaaacggaa acgggactct agcatactta ctgacagcca
gacagcgacc 2280aaaagaattc ggatggccat caattagtgt cgaatagttt
aaaaacgacc ttgtttctac 2340t 2341121565DNAInfluenza A virus
12agcaaaagca gggtagataa tcactcactg agtgacatca aaatcatggc gtctcaaggc
60accaaacgat cttacgaaca gatggagact gatggagaac gccagaatgc cactgaaatc
120agagcatccg tcggaaaaat gattggtgga attggacgat tctacatcca
aatgtgcacc 180gaactcaaac tcagtgatta tgagggacgg ttgatccaaa
acagcttaac aatagagaga 240atggtgctct ctgcttttga cgaaaggaga
aataaatacc ttgaagaaca tcccagtgcg 300ggaaaagatc ctaagaaaac
tggaggacct atatacagga gagtaaacgg aaagtggatg 360agagaactca
tcctttatga caaagaagaa ataaggcgaa tctggcgcca agctaataat
420ggtgacgatg caacggctgg tctgactcac atgatgatct ggcattccaa
tttgaatgat 480gcaacttatc agaggacaag agctcttgtt cgcaccggaa
tggatcccag gatgtgctct 540ctgatgcaag gttcaactct ccctaggagg
tctggagccg caggtgctgc agtcaaagga 600gttggaacaa tggtgatgga
attggtcaga atgatcaaac gtgggatcaa tgatcggaac 660ttctggaggg
gtgagaatgg acgaaaaaca agaattgctt atgaaagaat gtgcaacatt
720ctcaaaggga aatttcaaac tgctgcacaa aaagcaatga tggatcaagt
gagagagagc 780cggaacccag ggaatgctga gttcgaagat ctcacttttc
tagcacggtc tgcactcata 840ttgagagggt cggttgctca caagtcctgc
ctgcctgcct gtgtgtatgg acctgccgta 900gccagtgggt acgactttga
aagggaggga tactctctag tcggaataga ccctttcaga 960ctgcttcaaa
acagccaagt gtacagccta atcagaccaa atgagaatcc agcacacaag
1020agtcaactgg tgtggatggc atgccattct gccgcatttg aagatctaag
agtattaagc 1080ttcatcaaag ggacgaaggt gctcccaaga gggaagcttt
ccactagagg agttcaaatt 1140gcttccaatg aaaatatgga gactatggaa
tcaagtacac ttgaactgag aagcaggtac 1200tgggccataa ggaccagaag
tggaggaaac accaatcaac agagggcatc tgcgggccaa 1260atcagcatac
aacctacgtt ctcagtacag agaaatctcc cttttgacag aacaaccatt
1320atggcagcat tcaatgggaa tacagagggg agaacatctg acatgaggac
cgaaatcata 1380aggatgatgg aaagtgcaag accagaagat gtgtctttcc
aggggcgggg agtcttcgag 1440ctctcggacg aaaaggcagc gagcccgatc
gtgccttcct ttgacatgag taatgaagga 1500tcttatttct tcggagacaa
tgcagaggag tacgacaatt aaagaaaaat acccttgttt 1560ctact
1565131027DNAInfluenza A virus 13agcaaaagca ggtagatatt gaaagatgag
tcttctaacc gaggtcgaaa cgtacgtact 60ctctatcatc ccgtcaggcc ccctcaaagc
cgagatcgca cagagacttg aagatgtctt 120tgcagggaag aacaccgatc
ttgaggttct catggaatgg ctaaagacaa gaccaatcct 180gtcacctctg
actaagggga ttttaggatt tgtgttcacg ctcaccgtgc ccagtgagcg
240aggactgcag cgtagacgct ttgtccaaaa tgcccttaat gggaacgggg
atccaaataa 300catggacaaa gcagttaaac tgtataggaa gctcaagagg
gagataacat tccatggggc 360caaagaaatc tcactcagtt attctgctgg
tgcacttgcc agttgtatgg gcctcatata 420caacaggatg ggggctgtga
ccactgaagt ggcatttggc ctggtatgtg caacctgtga 480acagattgct
gactcccagc atcggtctca taggcaaatg gtgacaacaa ccaatccact
540aatcagacat gagaacagaa tggttttagc cagcactaca gctaaggcta
tggagcaaat 600ggctggatcg agtgagcaag cagcagaggc catggaggtt
gctagtcagg ctagacaaat 660ggtgcaagcg atgagaacca ttgggactca
tcctagctcc agtgctggtc tgaaaaatga 720tcttcttgaa aatttgcagg
cctatcagaa acgaatgggg gtgcagatgc aacggttcaa 780gtgatcctct
cactattgcc gcaaatatca ttgggatctt gcacttgaca ttgtggattc
840ttgatcgtct ttttttcaaa tgcatttacc gtcgctttaa atacggactg
aaaggagggc 900cttctacgga aggagtgcca aagtctatga gggaagaata
tcgaaaggaa cagcagagtg 960ctgtggatgc tgacgatggt cattttgtca
gcatagagct ggagtaaaaa actaccttgt 1020ttctact 102714890DNAInfluenza
A virus 14agcaaaagca gggtgacaaa aacataatgg atccaaacac tgtgtcaagc
tttcaggtag 60attgctttct ttggcatgtc cgcaaacgag ttgcagacca agaactaggt
gatgccccat 120tccttgatcg gcttcgccga gatcagaaat ccctaagagg
aaggggcagt actctcggtc 180tggacatcaa gacagccaca cgtgctggaa
agcagatagt ggagcggatt ctgaaagaag 240aatccgatga ggcacttaaa
atgaccatgg cctctgtacc tgcgtcgcgt tacctaactg 300acatgactct
tgaggaaatg tcaagggact ggtccatgct catacccaag cagaaagtgg
360caggccctct ttgtatcaga atggaccagg cgatcatgga taagaacatc
atactgaaag 420cgaacttcag tgtgattttt gaccggctgg agactctaat
attgctaagg gctttcaccg 480aagagggagc aattgttggc gaaatttcac
cattgccttc tcttccagga catactgctg 540aggatgtcaa aaatgcagtt
ggagtcctca tcggaggact tgaatggaat gataacacag 600ttcgagtctc
tgaaactcta cagagattcg cttggagaag cagtaatgag aatgggagac
660ctccactcac tccaaaacag aaacgagaaa tggcgggaac aattaggtca
gaagtttgaa 720gaaataagat ggttgattga agaagtgaga cacaaactga
agataacaga gaatagtttt 780gagcaaataa catttatgca agccttacat
ctattgcttg aagtggagca agagataaga 840actttctcgt ttcagcttat
ttagtactaa aaaacaccct tgtttctact 890151775DNAInfluenza A virus
15agcaaaagca ggggaaaata aaaacaacca aaatgaaggc aaacctactg gtcctgttat
60gtgcacttgc agctgcagat gcagacacaa tatgtatagg ctaccatacg aacaattcaa
120ccgacactgt tgacacagta ctcgagaaga atgtgacagt gacacactct
gttaacctgc 180tcgaagacag ccacaacgga aaactatgta gattaaaagg
aatagcccca ctacaattgg 240ggaaatgtaa catcgccgga tggctcttgg
gaaacccaga atgcgaccca ctgcttccag 300tgagatcatg gtcctacatt
gtagaaacac caaactctga gaatggaata tgttatccag 360gagatttcat
cgactatgag gagctgaggg agcaattgag ctcagtgtca tcattcgaaa
420gattcgaaat atttcccaaa gaaagctcat ggcccaacca caacacaaac
ggagtaacgg 480cagcatgctc ccatgagggg aaaagcagtt tttacagaaa
tttgctatgg ctgacggaga 540aggagggctc atacccaaag ctgaaaaatt
cttatgtgaa caaaaaaggg aaagaagtcc 600ttgtactgtg gggtattcat
cacccgccta acagtaagga acaacagaat ctctatcaga 660atgaaaatgc
ttatgtctct gtagtgactt caaattataa caggagattt accccggaaa
720tagcagaaag acccaaagta agagatcaag ctgggaggat gaactattac
tggaccttgc 780taaaacccgg agacacaata atatttgagg caaatggaaa
tctaatagca ccaatgtatg 840ctttcgcact gagtagaggc tttgggtccg
gcatcatcac ctcaaacgca tcaatgcatg 900agtgtaacac gaagtgtcaa
acacccctgg gagctataaa cagcagtctc ccttaccaga 960atatacaccc
agtcacaata ggagagtgcc caaaatacgt caggagtgcc aaattgagga
1020tggttacagg actaaggaac attccgtcca ttcaatccag aggtctattt
ggagccattg 1080ccggttttat tgaaggggga tggactggaa tgatagatgg
atggtatggt tatcatcatc 1140agaatgaaca gggatcaggc tatgcagcgg
atcaaaaaag cacacaaaat gccattaacg 1200ggattacaaa caaggtgaac
actgttatcg agaaaatgaa cattcaattc acagctgtgg 1260gtaaagaatt
caacaaatta gaaaaaagga tggaaaattt aaataaaaaa gttgatgatg
1320gatttctgga catttggaca tataatgcag aattgttagt tctactggaa
aatgaaagga 1380ctctggaatt ccatgactca aatgtgaaga atctgtatga
gaaagtaaaa agccaattaa 1440agaataatgc caaagaaatc ggaaatggat
gttttgagtt ctaccacaag tgtgacaatg 1500aatgcatgga aagtgtaaga
aatgggactt atgattatcc caaatattca gaagagtcaa 1560agttgaacag
ggaaaaggta gatggagtga aattggaatc aatggggatc tatcagattc
1620tggcgatcta ctcaactgtc gccagttcac tggtgctttt ggtctccctg
ggggcaatca 1680gtttctggat gtgttctaat ggatctttgc agtgcagaat
atgcatctga gattagaatt 1740tcagagatat gaggaaaaac acccttgttt ctact
1775161413DNAInfluenza A virus 16agcaaaagca ggggtttaaa atgaatccaa
atcagaaaat aataaccatt ggatcaatct 60gtctggtagt cggactaatt agcctaatat
tgcaaatagg gaatataatc tcaatatgga 120ttagccattc aattcaaact
ggaagtcaaa accatactgg aatatgcaac caaaacatca 180ttacctataa
aaatagcacc tgggtaaagg acacaacttc agtgatatta accggcaatt
240catctctttg tcccatccgt gggtgggcta tatacagcaa agacaatagc
ataagaattg 300gttccaaagg agacgttttt gtcataagag agccctttat
ttcatgttct cacttggaat 360gcaggacctt ttttctgacc caaggtgcct
tactgaatga caagcattca agtgggactg 420ttaaggacag aagcccttat
agggccttaa tgagctgccc tgtcggtgaa gctccgtccc 480cgtacaattc
aagatttgaa tcggttgctt ggtcagcaag tgcatgtcat gatggcatgg
540gctggctaac aatcggaatt tcaggtccag ataatggagc agtggctgta
ttaaaataca 600acggcataat aactgaaacc ataaaaagtt ggaggaagaa
aatattgagg acacaagagt 660ctgaatgtgc ctgtgtaaat ggttcatgtt
ttactataat gactgatggc ccgagtgatg 720ggctggcctc gtacaaaatt
ttcaagatcg aaaaggggaa ggttactaaa tcaatagagt 780tgaatgcacc
taattctcac tatgaggaat gttcctgtta ccctgatacc gacaaagtga
840tgtgtgtgtg cagagacaat tggcatggtt cgaaccggcc atgggtgtct
ttcgatcaaa 900acctggatta tcaaatagga tacatctgca gtggggtttt
cggtgacaac ccgcgtcccg 960aagatggaac aggcagctgt ggtccagtgt
atgttgatgg agcaaacgga gtaaagggat 1020tttcatatag gtatggtaat
ggtgtttgga taggaaggac caaaagtcac agttccagac 1080atgggtttga
gatgatttgg gatcctaatg gatggacaga gactgatagt aagttctctg
1140tgaggcaaga tgttgtggca atgactgatt ggtcagggta tagcggaagt
ttcgttcaac 1200atcctgagct gacagggcta gactgtatga ggccgtgctt
ctgggttgaa ttaatcaggg 1260gacgacctaa agaaaaaaca atctggacta
gtgcgagcag catttctttt tgtggcgtga 1320atagtgatac tgtagattgg
tcttggccag acggtgctga gttgccattc agcattgaca 1380agtagtctgt
tcaaaaaact ccttgtttct act 1413172220DNAInfluenza A virus
17agcgaaagca ggtactgatt cgaaatggaa gattttgtgc gacaatgctt caatccgatg
60attgtcgagc ttgcggaaaa ggcaatgaaa gagtatggag aggacctgaa aatcgaaaca
120aacaaatttg cagcaatatg cacccacttg gaagtatgct tcatgtattc
agattttcat 180ttcatcaatg agcaaggcga atcaataata gtagagcctg
aggacccaaa tgcactttta 240aaacacagat ttgagataat agaggggcga
gatcgtacaa tggcatggac agttgtaaac 300agtatttgca acaccacagg
agctgagaaa ccaaagtttc tgccagatct gtatgattac 360aaagagaata
ggttcatcga aattggagtg acaaggagag aagttcacat atactatctg
420gaaaaggcca acaaaattaa atctgagaag acacatattc acattttctc
atttactggc 480gaagaaatgg ccacaaaggc cgattacact ctcgatgaag
aaagcagggc tagaattaaa 540accagactat tcaccataag gcaagaaatg
gcaagcagag gtctttggga ctcctttcgt 600cagtccgaaa gaggcgaaga
gacaattgaa gaaaggtttg aaatcacagg gacaatgcgc 660aggctcgctg
atcaaagcct tccgccgaac ttctcctgca ttgagaattt tagagcctat
720gtggatggat ttgaaccgaa cggctacatt gagggcaagc tttctcaaat
gtccaaagaa 780gtaaatgcta aaattgagcc ttttttgaaa acaacacctc
gaccaattag acttccgaat 840gggcctcctt gttttcagcg gtcaaaattc
ctgctgatgg attctttaaa attaagcatt 900gaggatccaa atcatgaagg
ggagggaata ccactatatg atgcaatcaa gtgtatgaga 960acattctttg
gatggaaaga acccactgtt gtcaagccac acgagaaggg aataaatccg
1020aattatctgc tgtcgtggaa gcaggtgttg gaagagctgc aggacattga
gagtgaggag 1080aagattccaa gaacaaaaaa catgaaaaaa acgagtcagt
taaagtgggc acttggtgag 1140aacatggcac cagagaaggt ggattttgat
gactgtaaag atataagcga tttgaagcaa 1200tatgatagtg acgaacctga
attaaggtca ttttcaagtt ggatccagaa tgagttcaac 1260aaggcatgcg
agctgaccga ttcaatctgg atagagctcg atgagattgg agaagatgtg
1320gccccgattg aacacattgc aagcatgaga agaaattact tcacagctga
ggtgtcccat 1380tgcagagcca ctgaatatat aatgaaaggg gtatacatta
atactgcttt gcttaatgca 1440tcctgtgcag caatggatga tttccaacta
attcctatga taagcaaatg tagaactaaa 1500gagggaagga gaaagaccaa
tttgtacggc ttcatcataa aaggaagatc tcacttaagg 1560aatgataccg
atgtggtaaa ctttgtgagc atggagtttt ccctcactga cccaagactt
1620gagccacaca aatgggagaa gtactgtgtt cttgagatag gagatatgct
tctaaggagt 1680gcaataggcc aagtgtcaag gcccatgttc ttgtatgtaa
gaacaaatgg aacctcaaaa 1740attaaaatga aatggggaat ggagatgagg
cgttgcctcc tccaatccct ccaacaaata 1800gagagcatga ttgaagctga
gtcctctgtc aaggagaaag acatgacaaa agagtttttt 1860gagaatagat
cagaaacatg gcccattgga gagtcaccaa aaggagtgga agaaggttcc
1920attgggaaag tatgcaggac actattggct aaatcagtat tcaatagtct
gtatgcatct 1980ccacaattag aaggattttc agctgagtca agaaagttgc
tccttattgt tcaggctctt 2040agggacaatc tggaacctgg gacctttgat
cttgggggac tatatgaagc aattgaggag 2100tgcctgatta atgatccctg
ggttttgctt aatgcttctt ggttcaactc cttcctaaaa
2160catgcattga gatagctgag gcaatgctac tatttgttat ccatactgtc
caaaaaagta 2220182341DNAInfluenza A virus 18agcgaaagca ggcaaaccat
ttgaatggat gtcaatccga cattactttt cttaaaagtg 60ccagcacaaa atgctataag
cacaactttt ccttatactg gtgaccctcc ttacagccat 120ggaacaggaa
caggatacac catggataca gtcaacagga cacatcagta ctcagaaaga
180ggaagatgga cgaaaaatac cgaaactgga gcaccgcaac tcaacccaat
tgatgggcca 240ctaccagaag acaatgaacc aagtggctat gcccaaacag
attgtgtatt agaggcaatg 300gctttccttg aagaatccca tcctggtatt
tttgaaaact cttgtattga aacaatggag 360gttgttcagc aaacaagggt
ggacaaactg acacaaggca gacaaaccta tgactggact 420ctaaatagga
accagcctgc tgccacagca ttggcaaaca ccatagaagt attcagatca
480aatggcctca tagcaaatga atctggaagg ctaatagact tccttaaaga
tgtaatggag 540tcgatggaca gagacgaagt agaggtcaca actcattttc
aaagaaagag gagagtgaga 600gacaatgtaa ctaaaaaaat ggtgacccaa
agaacaatag gaaaaaagaa acataaatta 660gacaaaagaa gttacctaat
tagggcatta accctgaaca caatgaccaa agatgctgag 720agggggaaac
taaaacgcag agcaattgca accccaggaa tgcaaataag ggggtttgta
780tactttgttg agacactggc aagaagcata tgtgaaaagc ttgaacaatc
agggttgcca 840gttggaggaa atgagaagaa agcaaagtta gcaaatgttg
taaggaagat gatgaccaac 900tcccaggaca ctgaaatttc ttttaccatc
actggagata acacaaaatg gaacgaaaat 960caaaacccta gaatgttctt
ggccatgatc acatatataa ccaaagatca gcctgaatgg 1020ttcagaaata
ttctaagtat tgctccaata atgttttcaa acaaaatggc gagactaggt
1080agggggtata tgtttgaaag caagagtatg aaactgagaa cccaaatacc
tgcagagatg 1140ctagccaaca tagatttgaa atatttcaat gattcaacta
aaaagaaaat tgaaaaaatt 1200cgaccattat taatagatgg aactgcatca
ttgagtcctg gaatgatgat gggcatgttc 1260aatatgttaa gcaccgtctt
gggcgtttcc attctgaatc ttgggcaaaa aagatacacc 1320aagactactt
actggtggga tggtcttcaa tcgtctgatg attttgcttt gattgtgaat
1380gcacccaatt atgcaggaat tcaagctgga gttgacaggt tttatcgaac
ctgtaagctg 1440ctcggaatta atatgagcaa aaagaagtct tacataaaca
gaacaggtac ctttgaattc 1500acgagctttt tctatcgtta tgggtttgtt
gccaatttca gcatggagct tcctagtttt 1560ggggtgtctg gggtcaatga
atctgcagac atgagtattg gagtcactgt catcaaaaac 1620aatatgataa
acaatgacct tggcccagca actgctcaaa tggcccttca gttatttata
1680aaagattaca ggtacactta tcgatgccac agaggtgaca cacaaataca
aacccggaga 1740tcatttgaaa taaagaaact atgggaccaa acccgctcca
aagctgggct gttggtctct 1800gatggaggcc ccaatttata taacattagg
aatctacata ttcctgaagt ctgcttgaaa 1860tgggagttga tggatgagga
ttaccagggg cgtttatgca acccattgaa cccgtttgtc 1920agccataaag
agattgaatc agtgaacaat gcagtgataa tgccggcaca tggtccagcc
1980aaaaatatgg agtatgacgc tgttgcaaca acacactctt gggtccccaa
aagaaatcga 2040tccattttaa acacgagcca aagagggata cttgaagatg
agcaaatgta ccaaaggtgc 2100tgcaatttat ttgaaaaatt cttcccaagt
agctcataca gaagaccagt tggaatatcc 2160agtatggtag aggctatggt
ttcaagagcc cgaattgatg cacggattga tttcgaatct 2220ggaaggataa
agaaagagga attcgctgag atcatgaaga cctgttccac cattgaagac
2280ctcagacggc aaaaataggg aatttggctt gtccttcatg aaaaaatgcc
ttgtttctac 2340t 2341192341DNAInfluenza A virus 19agcgaaagca
ggtcaattat attcaatatg gaaagaataa aagagctaag gaatctgatg 60tcacaatctc
gcactcgcga gatacttacc aaaactactg tagaccacat ggccataata
120aagaaataca catcaggaag acaggagaaa aacccatcac ttaggatgaa
atggatgatg 180gcaatgaaat acccaattac agctgataaa aggataacgg
aaatgattcc tgaaagaaat 240gagcaaggac agacactatg gagtaaagtg
aatgatgccg gatcagaccg agtgatgata 300tcacccctag ctgtgacatg
gtggaacaga aatggaccag tggcaaacac tatccactat 360ccaaaaatct
acaaaactta ctttgaaaag gttgaaaggt taaaacatgg aacctttggc
420cctgtacact ttagaaacca agtcaaaata cgccgaagag tcgacataaa
tcctggtcat 480gcagacctca gcgccaagga ggcacaggat gtaattatgg
aagttgtttt ccctaatgaa 540gtgggagcca gaatactaac atcagaatcg
caattaacga taactaagga gaaaaaagag 600gaactccaga attgcaaaat
ttcccctttg atggttgcat acatgttaga gagggaactt 660gtccgcaaaa
caagatttct cccggttgca ggtggaacaa gcagtgtgta cattgaagtt
720ttgcatttaa cacaggggac atgctgggag cagatgtaca ctccaggtgg
ggaggtgagg 780aatgatgatg ttgatcaaag cctaattatt gctgctagga
acatagtgag aagagctgca 840gtatcagcag atccactagc atctttatta
gaaatgtgcc atagcacaca gattggtgga 900acaaggatgg tggatattct
caggcaaaat ccaacagaag aacaagctgt ggacatatgc 960aaagcagcaa
tggggctgag aatcagttca tccttcagtt ttggcggatt cacatttaag
1020agaacaagtg gatcgtcagt caaaagggag gaagaagtgc taacgggcaa
tctgcaaaca 1080ttgaagctaa ctgtgcatga gggatatgaa gaattcacaa
tagttgggaa aaaggcaaca 1140gctatactca gaaaagcaac caggagattg
attcaactaa tagtgagtgg aagagacgaa 1200cagtcaatag tcgaagcaat
agttgtagca atggtattct cacaagaaga ttgcatggta 1260aaagcggtta
gaggtgatct gaatttcgtt aatagagcga atcagcggtt gaatcccatg
1320catcaacttt tgagacattt tcagaaggat gctaaagtac ttttcctaaa
ttggggaatt 1380gaacatattg acaatgtgat gggaatgatt gggatattac
ctgatatgac tccaagtacc 1440gagatgtcaa tgagaggagt gagagtcagc
aaaatgggtg tagatgaata ctccaatgct 1500gaaagggtag tggtaagcat
tgaccgtttt ttgagggtcc gggaccaaag aggaaatgta 1560ttactgtctc
cagaggaagt cagtgaaaca caaggaacag agaaactgac aataacttac
1620tcttcatcat tgatgtggga gattaatggc cctgagtcag tgttgatcaa
tacctaccaa 1680tggatcatca gaaactggga gactgttaaa attcagtggt
ctcagaaccc tacaatgcta 1740tacaataaaa tggaatttga gccatttcaa
tctctagtcc ccaaggccat tagaggccaa 1800tacagtgggt ttgttagaac
tctatttcaa caaatgaggg atgtgctcgg gacctttgac 1860acaactcaga
taataaaact tcttcccttt gcagccgctc caccaaagca aagtagaatg
1920caattctcgt cattaactgt gaatgtgagg ggatcaggaa tgagaatact
tgtaaggggt 1980aattctccag tattcaacta caacaagacc actaagagac
tcacaatcct cggaaaggat 2040gctggcactt taactgaaga cccagatgaa
ggcacagctg gagtggaatc tgctgtttta 2100aggggattcc tcattctagg
caaagaagat agaagatatg ggccagcatt aagcatcagt 2160gaattgagca
accttgcgaa aggggagaaa gctaatgtgc taattgggca aggggatgta
2220gtgttggtaa tgaaacgaaa acgggactct agcatactta ctgacagcca
gacagcgacc 2280aaaagaattc ggatggccat caattaattt cgaataattt
aaaaacgacc ttgtttctac 2340t 2341201565DNAInfluenza A virus
20agcaaaagca gggtagataa tcactcactg agtgacatca aagtcatggc gtcccaaggc
60accaaacggt cttacgaaca gatggagact gatggggaac gccagaatgc aactgaaatc
120agagcatccg tcggaagaat gattggggga attgggcgat tctacatcca
aatgtgcacc 180gagcttaagc tcaatgatta tgagggacga ctgatccaga
acagcttaac aatagagaga 240atggtgcttt ctgcttttga tgagaggaga
aataaatatc tggaagaaca tcccagcgca 300gggaaagatc ctaagaaaac
tggaggaccc atatacaaga gagtagatgg aaagtgggtg 360agggaactcg
tcctttatga caaagaagaa ataaggcgga tttggcgcca agccaacaat
420ggtgatgatg caacagctgg tttgactcac attatgatct ggcattctaa
tttgaatgat 480acaacttacc agaggacaag agctcttgtc cgcaccggaa
tggatcccag gatgtgctct 540ttgatgcaag gttcaactct ccctagaaga
tctggagcag caggcgctgc agtcaaagga 600gttgggacaa tggtattgga
gttaatcagg atgatcaaac gtgggatcaa cgaccgaaac 660ttctggaggg
gtgagaatgg gagaaaaaca aggattgctt atgagagaat gtgcaacatt
720ctcaaaggaa aatttcaaac agctgcacaa aaagcaatga tggatcaagt
gagagaaagc 780cggaacccag gaaatgctga gatcgaagat ctcacttttc
tggcacggtc tgcactcata 840ttgagaggat cagttgctca caagtcttgc
ctgcctgctt gtgtgtatgg accagccgta 900gccagtgggt atgacttcga
aaaagaggga tactctttgg tgggagtaga ccctttcaaa 960ctgcttcaaa
ccagtcaggt atacagccta attagaccaa acgagaatcc cgcacacaag
1020agccagttgg tgtggatggc atgcaattct gctgcatttg aagatctaag
agtgtcaagc 1080ttcatcagag ggacaagagt acttccaagg gggaagctct
ccactagagg agtacaaatt 1140gcttcaaatg aaaacatgga tgctattgtc
tcaagtactc ttgaactgag aagcagatac 1200tgggccataa gaaccagaag
tggagggaac accaatcaac aaagggcctc tgcgggccaa 1260atcagcacac
aacctacgtt ttctgtgcag agaaacctcc catttgacaa aacaaccatc
1320atggcagcat tcactgggaa tacagaggga agaacatcag acatgcgggc
agaaatcata 1380aagatgatgg aaagtgcaag accagaagaa gtgtccttcc
agggacgggg agtctttgag 1440ctctcggacg aaagggcaac gaacccgatc
gtgccctcct ttgacatgag taatgaagga 1500tcttatttct tcggagacaa
tgcagaggag tacgacaatt aatgaaaaat acccttgttt 1560ctact
1565211027DNAInfluenza A virus 21agcaaaagca ggtagatatt gaaagatgag
tcttctaacc gaggtcgaaa cgtacgttct 60ctctatcgtc ccatcaggcc ccctcaaagc
cgagatcgca cagagacttg aagatgtatt 120tgctggaaag aataccgatc
ttgaggctct catggaatgg ctaaagacaa gaccaatcct 180gtcacctctg
actaagggga ttttaggatt tgtgttcacg ctcaccgtgc ccagtgagcg
240aggactgcag cgtagacgct ttgtccaaaa tgcccttaat gggaatgggg
atccaaataa 300tatggacaag gctgtcaaac tgtatcgaaa gcttaagagg
gagataacat tccatggggc 360caaagaaata gcactcagtt attctgctgg
agcacttgcc agttgtatgg gactcatata 420caacaggatg ggggctgtga
ccaccgaatc agcatttggc cttatatgtg caacctgtga 480acagattgcc
gactcccagc ataagtctca taggcaaatg gtaacaacaa ccaatccatt
540aataagacat gagaacagaa tggttctggc cagcactaca gctaaggcta
tggagcaaat 600ggctggatcg agtgaacaag cagctgaggc catggaggtt
gctagtcagg ccaggcagat 660ggtgcaggca atgagagcca ttgggactca
tcctagctct agcactggtc tgaaaaatga 720tctccttgaa aatttgcagg
cctatcagaa acgaatgggg gtgcagatgc aacgattcaa 780gtgatcctct
tgttgttgcc gcaagtataa ttgggattgt gcacctgata ttgtggatta
840ttgatcgcct tttttccaaa agcatttatc gtatttttaa acacggttta
aaaagagggc 900cttctacgga aggagtaccg gagtctatga gggaagaata
tcgagaggaa cagcagaatg 960ctgtggatgc tgacgatggt cattttgtca
gcatagagct agagtaaaaa actaccttgt 1020ttctact 102722889DNAInfluenza
A virus 22agcaaaagca gggtggcaaa gacataatgg attcccacac tgtgtcaagc
tttcaggtag 60attgtttcct ttggcatgtc cgcaaacaag ttgcagacca agatctaggc
gatgccccct 120tccttgatcg gcttcgccga gatcagaagt ctctaaaggg
acgaggcaac actctcggtc 180tgaacatcga aacagccact tgtgttggaa
agcaaatagt agagaggatt ctgaaagaag 240aatccgatga gacatttaga
atgaccatgg cctccgcact tgcttcgcgg tacctaactg 300acatgactgt
tgaagaaatg tcaagggact ggttcatgct catgcccaag cagaaagtgg
360ctggccctct ttgtgtcaga atggaccagg cgataatgga taagaacatc
atactgaaag 420cgaacttcag tgtgattttt gaccggttgg agaatctgac
attactaagg gctttcaccg 480aagagggagc aattgttggc gaaatttcac
cattgccttc ttttccagga catactaatg 540aggatgtcaa aaatgcaatt
ggggtcctca tcgggggact tgaatggaat gataacacag 600ttcgagtctc
tgaagctcta cagagattcg cttggagaag cagtaatgag actgggggac
660ctccattcac tacaacacag aaacggaaaa tggcgggaac aattaggtca
gaagtttgaa 720gaaataagat ggctgattga agaagtgagg cataaattga
agacgacaga gagtagtttt 780gaacaaataa catttatgca agcattacag
ctattgtttg aagtggaaca agagattaga 840acgttctcgt ttcagcttat
ttaatgataa aaacaccctt gtttctact 889231775DNAInfluenza A virus
23agcgaaagca ggggaaaata aaagcaacca aaatgaaagt aaaactactg gttctgttat
60gtacatttac agctacatat gcagacacaa tatgtatagg ctaccatgcc aacaactcaa
120ccgacactgt tgacacagta cttgagaaga atgtaacagt gacacactct
gtcaacctac 180ttgaggacag tcacaatgga aaactatgtc tactaaaagg
aatagcccca ctacaattgg 240gtaattgcag cgttgccgga tggatcttag
gaaacccaga atgcgaatta ctgatttcca 300aggaatcatg gtcctacatt
gtagaaacac caaatcctga gaatggaaca tgttacccag 360ggtatttcgc
cgactatgag gaactgaggg agcaattgag ttcagtatct tcatttgaaa
420ggttcgaaat attccccaaa gagagctcat ggcccaacca caccgtaacc
ggagtatcag 480catcatgctc ccataacggg aaaagcagtt tttacagaaa
tttgctatgg ctgacgggga 540agaatggttt gtacccaaac ctgagcaagt
cctatgcaaa caacaaagag aaagaagtcc 600ttgtactatg gggtgttcat
cacccgccta acatagggga ccaaagggcc ctctatcata 660cagaaaatgc
ttatgtctct gtagtgtctt cacattatag cagaagattc accccagaaa
720tagccaaaag acccaaggtg agagaccagg aaggaagaat caactactac
tggactctgc 780tggaacccgg ggatacaata atatttgagg caaatggaaa
tctaatagcg ccaaggtatg 840ctttcgcact gagtagaggc ttgggatcag
gaatcatcac ctcaaatgca ccaatggatg 900aatgtgatgc aaagtgtcaa
acacctcagg gagctataaa cagcagtctt cctttccaga 960atgtacaccc
agtcacaata ggagagtgtc caaagtatgt caggagtgca aaattaagga
1020tggttacagg actaaggaac atcccatcca ttcaatccag aggtttgttt
ggagcaattg 1080ccggtttcat tgaagggggg tggactggaa tggtagatgg
ttggtatggt tatcatcatc 1140agaatgagca aggatctggg tatgctgcag
atcaaaaaag cacacaaaat gccattaacg 1200ggattacaaa caaggtgaat
tctgtaattg agaaaatgaa cactcaattc acagctgtgg 1260gcaaagaatt
caacaaattg gaaagaagga tggaaaactt aaataaaaaa gttgatgatg
1320ggtttctaga catttggacc tataatgcag aattgttggt tctactggaa
aatgaaagga 1380ctttggattt ccatgactcc aacgtgaaga atctgtatga
gaaagtaaaa agccaattaa 1440agaataatgc caaagaaata ggaaacgggt
gttttgaatt ctatcacaag tgtaacgatg 1500aatgcatgga gagtgtgaaa
aatggaactt atgactatcc aaaatattcc gaagaatcaa 1560agttaaacag
agagaaaatt gatggagtga aattggaatc aatgggagtc tatcagattc
1620tggcgatcta ctcaacagtc gccagttccc tggttctttt ggtctccctg
ggggcaatca 1680gcttctggat gtgttccaat gggtctttgc agtgtagaat
atgcatctaa gaccagaatt 1740tcagaaatat aaggaaaaac acccttgttt ctact
1775241462DNAInfluenza A virus 24agcaaaagca ggagtttaaa atgaatccaa
atcaaaaaat aataaccatt ggatcaatca 60gtatagcaat cggaataatt agtctaatgt
tgcaaatagg aaatattatt tcaatatggg 120ctagtcactc aatccaaact
ggaagtcaaa accacactgg aatatgcaac caaaaaatca 180tcacatatga
aaacagcacc tgggtgaatc acacatatgt taatattaac aacactaatg
240ttgttgctgg aaaggacaaa acttcagtga cactggccgg caattcatct
ctttgtccta 300tcagtggatg ggctatatac acaaaagaca acagcataag
aattggctcc aaaggagatg 360tttttgtcat aagagaacct ttcatatcat
gttctcactt ggaatgcaga accttttttc 420tgacccaagg tgctctatta
aatgacaaac attcaaatgg aaccgttaag gacagaagtc 480cttatagggc
cttaatgagc tgtcctctag gtgaagcccc gtcaccatac aattcaaagt
540ttgaatcagt tgcatggtca gcaagcgcat gccatgatgg caagggctgg
ttaacaatcg 600gaatttctgg tccagacaat ggagctgtgg ctgtactaaa
atacaacgga ataataactg 660aaaccataaa aagttgggaa aagcgaatat
tgagaacaca agagtctgaa tgtgtttgtg 720tgaacgggtc atgtttcacc
ataatgaccg atggcccgag taatggggcc gcctcgtaca 780aaatcttcaa
gatcgaaaag gggaaggtta ctaaatcaac agagttgaat gcacccaatt
840ttcattatga ggaatgttcc tgttacccag acactggcac agtgatgtgt
gtatgcaggg 900acaactggca tggttcaaat cgaccttggg tatcttttaa
tcaaaacttg gattatcaaa 960taggatacat ctgcagtgga gtgttcggtg
acaatccgcg tcccaaagat gggaagggca 1020gctgtaatcc agtgactgtt
gatggagcag acggagttaa ggggttttca tacaaatatg 1080gtaatggtgt
ttggatagga aggactaaaa gtaacagact tagaaagggg tttgagatga
1140tttgggatcc taatggatgg acagataccg acagtgattt ctcagtgaaa
caggatgttg 1200tggcaataac tgattggtca gggtacagcg gaagtttcgt
ccaacatcct gagttaacag 1260gattggactg tataagacct tgcttctggg
ttgagttagt cagaggactg cctagagaaa 1320atacaacaat ctggactagt
gggagcagca tttctttttg tggcgttgat agtgatactg 1380caaattggtc
ttggccagac ggtgctgagt tgccgttcac cattgacaag tagctcgttg
1440aaaaaaactc cttgtttcta ct 146225566PRTInfluenza A virus 25Met
Lys Ala Lys Leu Leu Val Leu Leu Cys Ala Leu Ser Ala Thr Asp 1 5 10
15 Ala Asp Thr Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr
20 25 30 Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser
Val Asn 35 40 45 Leu Leu Glu Asp Asn His Asn Gly Lys Leu Cys Lys
Leu Lys Gly Ile 50 55 60 Ala Pro Leu Gln Leu Gly Lys Cys Ser Ile
Ala Gly Trp Ile Leu Gly 65 70 75 80 Asn Pro Glu Cys Glu Ser Leu Phe
Ser Lys Lys Ser Trp Ser Tyr Ile 85 90 95 Ala Glu Thr Pro Asn Ser
Glu Asn Gly Thr Cys Tyr Pro Gly Tyr Phe 100 105 110 Ala Asp Tyr Glu
Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe 115 120 125 Glu Arg
Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Lys His Asn 130 135 140
Val Thr Lys Gly Val Thr Ala Ala Cys Ser His Lys Gly Lys Ser Ser 145
150 155 160 Phe Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Asn Gly Ser
Tyr Pro 165 170 175 Asn Leu Ser Lys Ser Tyr Val Asn Asn Lys Glu Lys
Glu Val Leu Val 180 185 190 Leu Trp Gly Val His His Pro Ser Asn Ile
Glu Asp Gln Lys Thr Ile 195 200 205 Tyr Arg Lys Glu Asn Ala Tyr Val
Ser Val Val Ser Ser His Tyr Asn 210 215 220 Arg Arg Phe Thr Pro Glu
Ile Ala Lys Arg Pro Lys Val Arg Asn Gln 225 230 235 240 Glu Gly Arg
Ile Asn Tyr Tyr Trp Thr Leu Leu Glu Pro Gly Asp Thr 245 250 255 Ile
Ile Phe Glu Ala Asn Gly Asn Leu Ile Ala Pro Trp Tyr Ala Phe 260 265
270 Ala Leu Ser Arg Gly Phe Gly Ser Gly Ile Ile Thr Ser Asn Ala Ser
275 280 285 Met Asp Glu Cys Asp Ala Lys Cys Gln Thr Pro Gln Gly Ala
Ile Asn 290 295 300 Ser Ser Leu Pro Phe Gln Asn Val His Pro Val Thr
Ile Gly Glu Cys 305 310 315 320 Pro Lys Tyr Val Arg Ser Thr Lys Leu
Arg Met Val Thr Gly Leu Arg 325 330 335 Asn Ile Pro Ser Ile Gln Ser
Arg Gly Leu Phe Gly Ala Ile Ala Gly 340 345 350 Phe Ile Glu Gly Gly
Trp Thr Gly Met Ile Asp Gly Trp Tyr Gly Tyr 355 360 365 His His Gln
Asn Glu Gln Gly Ser Gly Tyr Ala Ala Asp Gln Lys Ser 370 375 380 Thr
Gln Asn Ala Ile Asn Gly Ile Thr Asn Lys Val Asn Ser Ile Ile 385 390
395 400 Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn
Lys 405 410 415 Leu Glu Lys Arg Met Glu Asn Leu Asn Lys Lys Val Asp
Asp Gly Phe 420 425 430 Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu
Val Leu Leu Glu Asn 435 440 445 Glu Arg Thr Leu Asp Phe His Asp Ser
Asn Val Lys Asn Leu Tyr Glu 450 455 460 Lys Val Lys Ser Gln Leu Lys
Asn Asn Ala Lys
Glu Ile Gly Asn Gly 465 470 475 480 Cys Phe Glu Phe Tyr His Lys Cys
Asn Asn Glu Cys Met Glu Ser Val 485 490 495 Lys Asn Gly Thr Tyr Asp
Tyr Pro Lys Tyr Ser Glu Glu Ser Lys Leu 500 505 510 Asn Arg Glu Lys
Ile Asp Gly Val Lys Leu Glu Ser Met Gly Val Tyr 515 520 525 Gln Ile
Leu Ala Ile Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Leu 530 535 540
Val Ser Leu Gly Ala Ile Ser Phe Trp Met Cys Ser Asn Gly Ser Leu 545
550 555 560 Gln Cys Arg Ile Cys Ile 565 26470PRTInfluenza A virus
26Met Asn Pro Asn Gln Lys Ile Ile Thr Ile Gly Ser Ile Cys Met Thr 1
5 10 15 Ile Gly Ile Ile Ser Leu Ile Leu Gln Ile Gly Asn Ile Ile Ser
Ile 20 25 30 Trp Val Ser His Ser Ile Gln Thr Gly Ser Gln Asn His
Thr Gly Ile 35 40 45 Cys Asn Gln Arg Ile Ile Thr Tyr Glu Asn Ser
Thr Trp Val Asn Gln 50 55 60 Thr Tyr Val Asn Ile Asn Asn Thr Asn
Val Val Ala Gly Lys Asp Thr 65 70 75 80 Thr Ser Val Thr Leu Ala Gly
Asn Ser Ser Leu Cys Pro Ile Arg Gly 85 90 95 Trp Ala Ile Tyr Ser
Lys Asp Asn Ser Ile Arg Ile Gly Ser Lys Gly 100 105 110 Asp Val Phe
Val Ile Arg Glu Pro Phe Ile Ser Cys Ser His Leu Glu 115 120 125 Cys
Arg Thr Phe Phe Leu Thr Gln Gly Ala Leu Leu Asn Asp Lys His 130 135
140 Ser Asn Gly Thr Val Lys Asp Arg Ser Pro Tyr Arg Ala Leu Met Ser
145 150 155 160 Cys Pro Ile Gly Glu Ala Pro Ser Pro Tyr Asn Ser Arg
Phe Glu Ser 165 170 175 Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly
Met Gly Trp Leu Thr 180 185 190 Ile Gly Ile Ser Gly Pro Asp Asp Gly
Ala Val Ala Val Leu Lys Tyr 195 200 205 Asn Gly Ile Ile Thr Glu Thr
Ile Lys Ser Trp Arg Lys Arg Ile Leu 210 215 220 Arg Thr Gln Glu Ser
Glu Cys Val Cys Val Asn Gly Ser Cys Phe Thr 225 230 235 240 Ile Met
Thr Asp Gly Pro Ser Asn Gly Pro Ala Ser Tyr Arg Ile Phe 245 250 255
Lys Ile Glu Lys Gly Lys Ile Thr Lys Ser Ile Glu Leu Asp Ala Pro 260
265 270 Asn Ser His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Thr Gly Thr
Val 275 280 285 Met Cys Val Cys Arg Asp Asn Trp His Gly Ser Asn Arg
Pro Trp Val 290 295 300 Ser Phe Asn Gln Asn Leu Asp Tyr Gln Ile Gly
Tyr Ile Cys Ser Gly 305 310 315 320 Val Phe Gly Asp Asn Pro Arg Pro
Lys Asp Gly Lys Gly Ser Cys Asp 325 330 335 Pro Val Thr Val Asp Gly
Ala Asp Gly Val Lys Gly Phe Ser Tyr Arg 340 345 350 Tyr Gly Asn Gly
Val Trp Ile Gly Arg Thr Lys Ser Asn Ser Ser Arg 355 360 365 Lys Gly
Phe Glu Met Ile Trp Asp Pro Asn Gly Trp Thr Asp Thr Asp 370 375 380
Ser Asn Phe Leu Val Lys Gln Asp Val Val Ala Met Thr Asp Trp Ser 385
390 395 400 Gly Tyr Ser Gly Ser Phe Val Gln His Pro Glu Leu Thr Gly
Leu Asp 405 410 415 Cys Met Arg Pro Cys Phe Trp Val Glu Leu Val Arg
Gly Arg Pro Arg 420 425 430 Glu Gly Thr Thr Val Trp Thr Ser Gly Ser
Ser Ile Ser Phe Cys Gly 435 440 445 Val Asn Ser Asp Thr Ala Asn Trp
Ser Trp Pro Asp Gly Ala Glu Leu 450 455 460 Pro Phe Thr Ile Asp Lys
465 470 27469PRTInfluenza A virus 27Met Asn Pro Asn Gln Lys Ile Ile
Thr Ile Gly Ser Val Cys Met Thr 1 5 10 15 Ile Gly Met Ala Asn Leu
Ile Leu Gln Ile Gly Asn Ile Ile Ser Ile 20 25 30 Trp Ile Ser His
Ser Ile Gln Leu Gly Asn Gln Asn Gln Ile Glu Thr 35 40 45 Cys Asn
Gln Ser Val Ile Thr Tyr Glu Asn Asn Thr Trp Val Asn Gln 50 55 60
Thr Tyr Val Asn Ile Ser Asn Thr Asn Phe Ala Ala Gly Gln Ser Val 65
70 75 80 Val Ser Val Lys Leu Ala Gly Asn Ser Ser Leu Cys Pro Val
Ser Gly 85 90 95 Trp Ala Ile Tyr Ser Lys Asp Asn Ser Val Arg Ile
Gly Ser Lys Gly 100 105 110 Asp Val Phe Val Ile Arg Glu Pro Phe Ile
Ser Cys Ser Pro Leu Glu 115 120 125 Cys Arg Thr Phe Phe Leu Thr Gln
Gly Ala Leu Leu Asn Asp Lys His 130 135 140 Ser Asn Gly Thr Ile Lys
Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160 Cys Pro Ile
Gly Glu Val Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175 Val
Ala Trp Ser Ala Ser Ala Cys His Asp Gly Ile Asn Trp Leu Thr 180 185
190 Ile Gly Ile Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr
195 200 205 Asn Gly Ile Ile Thr Asp Thr Ile Lys Ser Trp Arg Asn Asn
Ile Leu 210 215 220 Arg Thr Gln Glu Ser Glu Cys Ala Cys Val Asn Gly
Ser Cys Phe Thr 225 230 235 240 Val Met Thr Asp Gly Pro Ser Asn Gly
Gln Ala Ser Tyr Lys Ile Phe 245 250 255 Arg Ile Glu Lys Gly Lys Ile
Val Lys Ser Val Glu Met Asn Ala Pro 260 265 270 Asn Tyr His Tyr Glu
Glu Cys Ser Cys Tyr Pro Asp Ser Ser Glu Ile 275 280 285 Thr Cys Val
Cys Arg Asp Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300 Ser
Phe Asn Gln Asn Leu Glu Tyr Gln Ile Gly Tyr Ile Cys Ser Gly 305 310
315 320 Ile Phe Gly Asp Asn Pro Arg Pro Asn Asp Lys Thr Gly Ser Cys
Gly 325 330 335 Pro Val Ser Ser Asn Gly Ala Asn Gly Val Lys Gly Phe
Ser Phe Lys 340 345 350 Tyr Gly Asn Gly Val Trp Ile Gly Arg Thr Lys
Ser Ile Ser Ser Arg 355 360 365 Asn Gly Phe Glu Met Ile Trp Asp Pro
Asn Gly Trp Thr Gly Thr Asp 370 375 380 Asn Asn Phe Ser Ile Lys Gln
Asp Ile Val Gly Ile Asn Glu Trp Ser 385 390 395 400 Gly Tyr Ser Gly
Ser Phe Val Gln His Pro Glu Leu Thr Gly Leu Asp 405 410 415 Cys Ile
Arg Pro Cys Phe Trp Val Glu Leu Ile Arg Gly Arg Pro Lys 420 425 430
Glu Asn Thr Ile Trp Thr Ser Gly Ser Ser Ile Ser Phe Cys Gly Val 435
440 445 Asn Ser Asp Thr Val Gly Trp Ser Trp Pro Asp Gly Ala Glu Leu
Pro 450 455 460 Phe Thr Ile Asp Lys 465 282277DNAInfluenza A virus
28atggaacgca ttaaagaact gcgcaacctg atgagccaga gccgcacccg cgaaattctg
60accaaaacca ccgtggatca tatggcgatt attaaaaaat ataccagcgg ccgccaggaa
120aaaaacccga gcctgcgcat gaaatggatg atggcgatga aatatccgat
taccgcggat 180aaacgcatta ccgaaatgat tccggaacgc aacgaacagg
gccagaccct gtggagcaaa 240gtgaacgatg cgggcagcga tcgcgtgatg
attagcccgc tggcggtgac ctggtggaac 300cgcaacggcc cggtggcgag
caccattcat tatccgaaaa tttataaaac ctattttgaa 360aaagtggaac
gcctgaaaca tggcaccttt ggcccggtgc attttcgcaa ccaggtgaaa
420attcgccgcc gcgtggatat taacccgggc catgcggatc tgagcgcgaa
agaagcgcag 480gatgtgatta tggaagtggt gtttccgaac gaagtgggcg
cgcgcattct gaccagcgaa 540agccagctga ccattaccaa agaaaaaaaa
gaagaactgc agaactgcaa aattagcccg 600ctgatggtgg cgtatatgct
ggaacgcgaa ctggtgcgca aaacccgctt tctgccggtg 660gcgggcggca
ccagcagcgt gtatattgaa gtgctgcatc tgacccaggg cacctgctgg
720gaacagatgt ataccccggg cggcgaagtg cgcaacgatg atgtggatca
gagcctgatt 780attgcggcgc gcaacattgt gcgccgcgcg gcggtgagcg
cggatccgct ggcgagcctg 840ctggaaatgt gccatagcac ccagattggc
ggcacccgca tggtggatat tctgcgccag 900aacccgaccg aagaacaggc
ggtggatatt tgcaaagcgg cgatgggcct gcgcattagc 960agcagcttta
gctttggcgg ctttaccttt aaacgcacca gcggcagcag cgtgaaacgc
1020gaagaagaag tgctgaccgg caacctgcag accctgaaac tgaccgtgca
tgaaggctat 1080gaagaattta ccatggtggg caaacgcgcg accgcgattc
tgcgcaaagc gacccgccgc 1140ctgattcagc tgattgtgag cggccgcgat
gaacagagca ttgtggaagc gattgtggtg 1200gcgatggtgt ttagccagga
agattgcatg gtgaaagcgg tgcgcggcga tctgaacttt 1260gtgaaccgcg
cgaaccagcg cctgaacccg atgcatcagc tgctgcgcca ttttcagaaa
1320gatgcgaaag tgctgtttct gaactggggc attgaaccga ttgataacgt
gatgggcatg 1380attggcattc tgccggatat gaccccgagc accgaaatga
gcatgcgcgg cgtgcgcgtg 1440agcaaaatgg gcgtggatga atatagcaac
gcggaacgcg tggtggtgag cattgatcgc 1500tttctgcgcg tgcgcgatca
gcgcggcaac gtgctgctga gcccggaaga agtgagcgaa 1560acccagggca
ccgaaaaact gaccattacc tatagcagca gcatgatgtg ggaaattaac
1620ggcccggaaa gcgtgctgat taacacctat cagtggatta ttcgcaactg
ggaaaccgtg 1680aaaattcagt ggagccagaa cccgaccatg ctgtataaca
aaatggaatt tgaaccgttt 1740cagagcctgg tgccgaaagc gattcgcggc
cagtatagcg gctttgtgcg caccctgttt 1800cagcagatgc gcgatgtgct
gggcaccttt gataccaccc agattattaa actgctgccg 1860tttgcggcgg
cgccgccgaa acagagccgc atgcagttta gcagcctgac cgtgaacgtg
1920cgcggcagcg gcatgcgcat tctggtgcgc ggcaacagcc cggtgtttaa
ctataacaaa 1980accaccaaac gcctgaccgt gctgggcaaa gatgcgggca
ccctgaccga agatccggat 2040gaaggcaccg cgggcgtgga aagcgcggtg
ctgcgcggct ttctgattct gggcaaagaa 2100gatcgccgct atggcccggc
gctgagcatt aacgaactga gcaacctggc gaaaggcgaa 2160aaagcgaacg
tgctgattgg ccagggcgat gtggtgctgg tgatgaaacg caaacgcgat
2220agcagcattc tgaccgatag ccagaccgcg accaaacgca ttcgcatggc gattaac
227729716PRTInfluenza A virus 29Met Glu Asp Phe Val Arg Gln Cys Phe
Asn Pro Met Ile Val Glu Leu 1 5 10 15 Ala Glu Lys Ala Met Lys Glu
Tyr Gly Glu Asp Pro Lys Ile Glu Thr 20 25 30 Asn Lys Phe Ala Ala
Ile Cys Thr His Leu Glu Val Cys Phe Met Tyr 35 40 45 Ser Asp Phe
His Phe Ile Asp Glu Arg Gly Glu Ser Ile Ile Val Glu 50 55 60 Ser
Gly Asp Pro Asn Ala Leu Leu Lys His Arg Phe Glu Ile Ile Glu 65 70
75 80 Gly Arg Asp Arg Ile Met Ala Trp Thr Val Val Asn Ser Ile Cys
Asn 85 90 95 Thr Thr Gly Val Glu Lys Pro Lys Phe Leu Pro Asp Leu
Tyr Asp Tyr 100 105 110 Lys Glu Asn Arg Phe Ile Glu Ile Gly Val Thr
Arg Arg Glu Val His 115 120 125 Ile Tyr Tyr Leu Glu Lys Ala Asn Lys
Ile Lys Ser Glu Lys Thr His 130 135 140 Ile His Ile Phe Ser Phe Thr
Gly Glu Glu Met Ala Thr Lys Ala Asp 145 150 155 160 Tyr Thr Leu Asp
Glu Glu Ser Arg Ala Arg Ile Lys Thr Arg Leu Phe 165 170 175 Thr Ile
Arg Gln Glu Met Ala Ser Arg Ser Leu Trp Asp Ser Phe Arg 180 185 190
Gln Ser Glu Arg Gly Glu Glu Thr Ile Glu Glu Lys Phe Glu Ile Thr 195
200 205 Gly Thr Met Arg Lys Leu Ala Asp Gln Ser Leu Pro Pro Asn Phe
Pro 210 215 220 Ser Leu Glu Asn Phe Arg Ala Tyr Val Asp Gly Phe Glu
Pro Asn Gly 225 230 235 240 Cys Ile Glu Gly Lys Leu Ser Gln Met Ser
Lys Glu Val Asn Ala Lys 245 250 255 Ile Glu Pro Phe Leu Arg Thr Thr
Pro Arg Pro Leu Arg Leu Pro Asp 260 265 270 Gly Pro Leu Cys His Gln
Arg Ser Lys Phe Leu Leu Met Asp Ala Leu 275 280 285 Lys Leu Ser Ile
Glu Asp Pro Ser His Glu Gly Glu Gly Ile Pro Leu 290 295 300 Tyr Asp
Ala Ile Lys Cys Met Lys Thr Phe Phe Gly Trp Lys Glu Pro 305 310 315
320 Asn Ile Val Lys Pro His Glu Lys Gly Ile Asn Pro Asn Tyr Leu Met
325 330 335 Ala Trp Lys Gln Val Leu Ala Glu Leu Gln Asp Ile Glu Asn
Glu Glu 340 345 350 Lys Ile Pro Arg Thr Lys Asn Met Lys Arg Thr Ser
Gln Leu Lys Trp 355 360 365 Ala Leu Gly Glu Asn Met Ala Pro Glu Lys
Val Asp Phe Asp Asp Cys 370 375 380 Lys Asp Val Gly Asp Leu Lys Gln
Tyr Asp Ser Asp Glu Pro Glu Pro 385 390 395 400 Arg Ser Leu Ala Ser
Trp Val Gln Asn Glu Phe Asn Lys Ala Cys Glu 405 410 415 Leu Thr Asp
Ser Ser Trp Ile Glu Leu Asp Glu Ile Gly Glu Asp Val 420 425 430 Ala
Pro Ile Glu His Ile Ala Ser Met Arg Arg Asn Tyr Phe Thr Ala 435 440
445 Glu Val Ser His Cys Arg Ala Thr Glu Tyr Ile Met Lys Gly Val Tyr
450 455 460 Ile Asn Thr Ala Leu Leu Asn Ala Ser Cys Ala Ala Met Asp
Asp Phe 465 470 475 480 Gln Leu Ile Pro Met Ile Ser Lys Cys Arg Thr
Lys Glu Gly Arg Arg 485 490 495 Lys Thr Asn Leu Tyr Gly Phe Ile Ile
Lys Gly Arg Ser His Leu Arg 500 505 510 Asn Asp Thr Asp Val Val Asn
Phe Val Ser Met Glu Phe Ser Leu Thr 515 520 525 Asp Pro Arg Leu Glu
Pro His Lys Trp Glu Lys Tyr Cys Val Leu Glu 530 535 540 Ile Gly Asp
Met Leu Leu Arg Thr Ala Ile Gly Gln Val Ser Arg Pro 545 550 555 560
Met Phe Leu Tyr Val Arg Thr Asn Gly Thr Ser Lys Ile Lys Met Lys 565
570 575 Trp Gly Met Glu Met Arg Arg Cys Leu Leu Gln Ser Leu Gln Gln
Ile 580 585 590 Glu Ser Met Ile Glu Ala Glu Ser Ser Val Lys Glu Lys
Asp Met Thr 595 600 605 Lys Glu Phe Phe Glu Asn Lys Ser Glu Thr Trp
Pro Ile Gly Glu Ser 610 615 620 Pro Arg Gly Val Glu Glu Gly Ser Ile
Gly Lys Val Cys Arg Thr Leu 625 630 635 640 Leu Ala Lys Ser Val Phe
Asn Ser Leu Tyr Ala Ser Pro Gln Leu Glu 645 650 655 Gly Phe Ser Ala
Glu Ser Arg Lys Leu Leu Leu Ile Val Gln Ala Leu 660 665 670 Arg Asp
Asn Leu Glu Pro Gly Thr Phe Asp Leu Gly Gly Leu Tyr Glu 675 680 685
Ala Ile Glu Glu Cys Leu Ile Asn Asp Pro Trp Val Leu Leu Asn Ala 690
695 700 Ser Trp Phe Asn Ser Phe Leu Thr His Ala Leu Lys 705 710 715
30757PRTInfluenza A virus 30Met Asp Val Asn Pro Thr Leu Leu Phe Leu
Lys Val Pro Ala Gln Asn 1 5 10 15 Ala Ile Ser Thr Thr Phe Pro Tyr
Thr Gly Asp Pro Pro Tyr Ser His 20 25 30 Gly Thr Gly Thr Gly Tyr
Thr Met Asp Thr Val Asn Arg Thr His Gln 35 40 45 Tyr Ser Glu Arg
Gly Arg Trp Thr Lys Asn Thr Glu Thr Gly Ala Pro 50 55 60 Gln Leu
Asn Pro Ile Asp Gly Pro Leu Pro Lys Asp Asn Glu Pro Ser 65 70 75 80
Gly Tyr Ala Gln Thr Asp Cys Val Leu Glu Ala Met Ala Phe Leu Glu 85
90 95 Glu Ser His Pro Gly Ile Phe Glu Asn Ser Cys Ile Glu Thr Met
Glu 100 105 110 Val Val Gln Gln Thr Arg Val Asp Lys Leu Thr Gln Gly
Arg Gln Thr 115 120 125 Tyr Asp Trp Thr Leu Asn Arg Asn Gln Pro Ala
Ala Thr Ala Leu Ala 130 135 140 Asn Thr Ile Glu Val Phe Arg Ser Asn
Gly Leu Ile Ala Asn Glu Ser 145 150 155 160 Gly Arg Leu Ile Asp Phe
Leu Lys Asp Val Met Glu Ser Met Asp Arg 165 170 175 Asp Glu Val Glu
Val Thr Thr His Phe Gln Arg Lys Arg Arg Val Arg 180
185 190 Asp Asn Val Thr Lys Lys Met Val Thr Gln Arg Thr Ile Gly Lys
Lys 195 200 205 Lys His Lys Leu Asp Lys Arg Ser Tyr Leu Ile Arg Ala
Leu Thr Leu 210 215 220 Asn Thr Met Thr Lys Asp Ala Glu Arg Gly Lys
Leu Lys Arg Arg Ala 225 230 235 240 Ile Ala Thr Pro Gly Met Gln Ile
Arg Gly Phe Val Tyr Phe Val Glu 245 250 255 Thr Leu Ala Arg Ser Ile
Cys Glu Lys Leu Glu Gln Ser Gly Leu Pro 260 265 270 Val Gly Gly Asn
Glu Lys Lys Ala Lys Leu Ala Asn Val Val Arg Lys 275 280 285 Met Met
Thr Asn Ser Gln Asp Thr Glu Ile Ser Phe Thr Ile Thr Gly 290 295 300
Asp Asn Thr Lys Trp Asn Glu Asn Gln Asn Pro Arg Met Phe Leu Ala 305
310 315 320 Met Ile Thr Tyr Ile Thr Lys Asn Gln Pro Glu Trp Phe Arg
Asn Ile 325 330 335 Leu Ser Ile Ala Pro Ile Met Phe Ser Asn Lys Met
Ala Arg Leu Gly 340 345 350 Lys Gly Tyr Met Phe Glu Ser Lys Ser Met
Lys Leu Arg Thr Gln Ile 355 360 365 Pro Ala Glu Met Leu Ala Asn Ile
Asp Leu Lys Tyr Phe Asn Asp Ser 370 375 380 Thr Lys Arg Lys Ile Glu
Lys Ile Arg Pro Leu Leu Ile Asp Gly Thr 385 390 395 400 Ala Ser Leu
Ser Pro Gly Met Met Met Gly Met Phe Asn Met Leu Ser 405 410 415 Thr
Val Leu Gly Val Ser Ile Leu Asn Leu Gly Gln Lys Arg Tyr Thr 420 425
430 Lys Thr Thr Tyr Trp Trp Asp Gly Leu Gln Ser Ser Asp Asp Phe Ala
435 440 445 Leu Ile Val Asn Ala Pro Asn Tyr Ala Gly Ile Gln Ala Gly
Val Asp 450 455 460 Arg Phe Tyr Arg Thr Cys Lys Leu Leu Gly Ile Asn
Met Ser Lys Lys 465 470 475 480 Lys Ser Tyr Ile Asn Arg Thr Gly Thr
Phe Glu Phe Thr Ser Phe Phe 485 490 495 Tyr Arg Tyr Gly Phe Val Ala
Asn Phe Ser Met Glu Leu Pro Ser Phe 500 505 510 Gly Val Ser Gly Val
Asn Glu Ser Ala Asp Met Ser Ile Gly Val Thr 515 520 525 Val Ile Lys
Asn Asn Met Ile Asn Asn Asp Leu Gly Pro Ala Thr Ala 530 535 540 Gln
Met Ala Leu Gln Leu Phe Ile Lys Asp Tyr Arg Tyr Thr Tyr Arg 545 550
555 560 Cys His Arg Gly Asp Thr Gln Ile Gln Thr Arg Arg Ser Phe Glu
Ile 565 570 575 Lys Lys Leu Trp Asp Gln Thr Arg Ser Lys Ala Gly Leu
Leu Val Ser 580 585 590 Asp Gly Gly Pro Asn Leu Tyr Asn Ile Arg Asn
Leu His Ile Pro Glu 595 600 605 Val Cys Leu Lys Trp Glu Leu Met Asp
Glu Asp Tyr Gln Gly Arg Leu 610 615 620 Cys Asn Pro Ser Asn Pro Phe
Val Ser His Lys Glu Ile Glu Ser Val 625 630 635 640 Asn Asn Ala Val
Met Met Pro Ala His Gly Pro Ala Lys Asn Met Glu 645 650 655 Tyr Asp
Ala Val Ala Thr Thr His Ser Trp Val Pro Lys Arg Asn Arg 660 665 670
Ser Ile Leu Asn Thr Ser Gln Arg Gly Ile Leu Glu Asp Glu Gln Met 675
680 685 Tyr Gln Arg Cys Cys Asn Leu Phe Glu Lys Phe Phe Pro Ser Ser
Ser 690 695 700 Tyr Arg Arg Pro Val Gly Ile Ser Ser Met Val Glu Ala
Met Val Ser 705 710 715 720 Arg Ala Arg Ile Asp Ala Arg Ile Asp Phe
Glu Ser Gly Arg Ile Lys 725 730 735 Lys Glu Glu Phe Ala Glu Ile Met
Lys Thr Cys Ser Thr Ile Glu Asp 740 745 750 Leu Arg Arg Gln Lys 755
31759PRTInfluenza A virus 31Met Glu Arg Ile Lys Glu Leu Arg Asn Leu
Met Ser Gln Ser Arg Thr 1 5 10 15 Arg Glu Ile Leu Thr Lys Thr Thr
Val Asp His Met Ala Ile Ile Lys 20 25 30 Lys Tyr Thr Ser Gly Arg
Gln Glu Lys Asn Pro Ser Leu Arg Met Lys 35 40 45 Trp Met Met Ala
Met Lys Tyr Pro Ile Thr Ala Asp Lys Arg Ile Thr 50 55 60 Glu Met
Ile Pro Glu Arg Asn Glu Gln Gly Gln Thr Leu Trp Ser Lys 65 70 75 80
Val Asn Asp Ala Gly Ser Asp Arg Val Met Ile Ser Pro Leu Ala Val 85
90 95 Thr Trp Trp Asn Arg Asn Gly Pro Val Ala Ser Thr Ile His Tyr
Pro 100 105 110 Lys Ile Tyr Lys Thr Tyr Phe Glu Lys Val Glu Arg Leu
Lys His Gly 115 120 125 Thr Phe Gly Pro Val His Phe Arg Asn Gln Val
Lys Ile Arg Arg Arg 130 135 140 Val Asp Ile Asn Pro Gly His Ala Asp
Leu Ser Ala Lys Glu Ala Gln 145 150 155 160 Asp Val Ile Met Glu Val
Val Phe Pro Asn Glu Val Gly Ala Arg Ile 165 170 175 Leu Thr Ser Glu
Ser Gln Leu Thr Ile Thr Lys Glu Lys Lys Glu Glu 180 185 190 Leu Gln
Asn Cys Lys Ile Ser Pro Leu Met Val Ala Tyr Met Leu Glu 195 200 205
Arg Glu Leu Val Arg Lys Thr Arg Phe Leu Pro Val Ala Gly Gly Thr 210
215 220 Ser Ser Val Tyr Ile Glu Val Leu His Leu Thr Gln Gly Thr Cys
Trp 225 230 235 240 Glu Gln Met Tyr Thr Pro Gly Gly Glu Val Arg Asn
Asp Asp Val Asp 245 250 255 Gln Ser Leu Ile Ile Ala Ala Arg Asn Ile
Val Arg Arg Ala Ala Val 260 265 270 Ser Ala Asp Pro Leu Ala Ser Leu
Leu Glu Met Cys His Ser Thr Gln 275 280 285 Ile Gly Gly Thr Arg Met
Val Asp Ile Leu Arg Gln Asn Pro Thr Glu 290 295 300 Glu Gln Ala Val
Asp Ile Cys Lys Ala Ala Met Gly Leu Arg Ile Ser 305 310 315 320 Ser
Ser Phe Ser Phe Gly Gly Phe Thr Phe Lys Arg Thr Ser Gly Ser 325 330
335 Ser Val Lys Arg Glu Glu Glu Val Leu Thr Gly Asn Leu Gln Thr Leu
340 345 350 Lys Leu Thr Val His Glu Gly Tyr Glu Glu Phe Thr Met Val
Gly Lys 355 360 365 Arg Ala Thr Ala Ile Leu Arg Lys Ala Thr Arg Arg
Leu Ile Gln Leu 370 375 380 Ile Val Ser Gly Arg Asp Glu Gln Ser Ile
Val Glu Ala Ile Val Val 385 390 395 400 Ala Met Val Phe Ser Gln Glu
Asp Cys Met Val Lys Ala Val Arg Gly 405 410 415 Asp Leu Asn Phe Val
Asn Arg Ala Asn Gln Arg Leu Asn Pro Met His 420 425 430 Gln Leu Leu
Arg His Phe Gln Lys Asp Ala Lys Val Leu Phe Leu Asn 435 440 445 Trp
Gly Ile Glu Pro Ile Asp Asn Val Met Gly Met Ile Gly Ile Leu 450 455
460 Pro Asp Met Thr Pro Ser Thr Glu Met Ser Met Arg Gly Val Arg Val
465 470 475 480 Ser Lys Met Gly Val Asp Glu Tyr Ser Asn Ala Glu Arg
Val Val Val 485 490 495 Ser Ile Asp Arg Phe Leu Arg Val Arg Asp Gln
Arg Gly Asn Val Leu 500 505 510 Leu Ser Pro Glu Glu Val Ser Glu Thr
Gln Gly Thr Glu Lys Leu Thr 515 520 525 Ile Thr Tyr Ser Ser Ser Met
Met Trp Glu Ile Asn Gly Pro Glu Ser 530 535 540 Val Leu Ile Asn Thr
Tyr Gln Trp Ile Ile Arg Asn Trp Glu Thr Val 545 550 555 560 Lys Ile
Gln Trp Ser Gln Asn Pro Thr Met Leu Tyr Asn Lys Met Glu 565 570 575
Phe Glu Pro Phe Gln Ser Leu Val Pro Lys Ala Ile Arg Gly Gln Tyr 580
585 590 Ser Gly Phe Val Arg Thr Leu Phe Gln Gln Met Arg Asp Val Leu
Gly 595 600 605 Thr Phe Asp Thr Thr Gln Ile Ile Lys Leu Leu Pro Phe
Ala Ala Ala 610 615 620 Pro Pro Lys Gln Ser Arg Met Gln Phe Ser Ser
Leu Thr Val Asn Val 625 630 635 640 Arg Gly Ser Gly Met Arg Ile Leu
Val Arg Gly Asn Ser Pro Val Phe 645 650 655 Asn Tyr Asn Lys Thr Thr
Lys Arg Leu Thr Val Leu Gly Lys Asp Ala 660 665 670 Gly Thr Leu Thr
Glu Asp Pro Asp Glu Gly Thr Ala Gly Val Glu Ser 675 680 685 Ala Val
Leu Arg Gly Phe Leu Ile Leu Gly Lys Glu Asp Arg Arg Tyr 690 695 700
Gly Pro Ala Leu Ser Ile Asn Glu Leu Ser Asn Leu Ala Lys Gly Glu 705
710 715 720 Lys Ala Asn Val Leu Ile Gly Gln Gly Asp Val Val Leu Val
Met Lys 725 730 735 Arg Lys Arg Asp Ser Ser Ile Leu Thr Asp Ser Gln
Thr Ala Thr Lys 740 745 750 Arg Ile Arg Met Ala Ile Asn 755
32498PRTInfluenza A virus 32Met Ala Ser Gln Gly Thr Lys Arg Ser Tyr
Glu Gln Met Glu Thr Asp 1 5 10 15 Gly Glu Arg Gln Asn Ala Thr Glu
Ile Arg Ala Ser Val Gly Arg Met 20 25 30 Ile Gly Gly Ile Gly Arg
Phe Tyr Ile Gln Met Cys Thr Glu Leu Lys 35 40 45 Leu Asn Asp Tyr
Glu Gly Arg Leu Ile Gln Asn Ser Leu Thr Ile Glu 50 55 60 Arg Met
Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu 65 70 75 80
Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro Ile 85
90 95 Tyr Lys Arg Val Asp Gly Lys Trp Val Arg Glu Leu Val Leu Tyr
Asp 100 105 110 Lys Glu Glu Ile Arg Arg Ile Trp Arg Gln Ala Asn Asn
Gly Asp Asp 115 120 125 Ala Thr Ala Gly Leu Thr His Ile Met Ile Trp
His Ser Asn Leu Asn 130 135 140 Asp Thr Thr Tyr Gln Arg Thr Arg Ala
Leu Val Arg Thr Gly Met Asp 145 150 155 160 Pro Arg Met Cys Ser Leu
Met Gln Gly Ser Thr Leu Pro Arg Arg Ser 165 170 175 Gly Ala Ala Gly
Ala Ala Val Lys Gly Val Gly Thr Met Val Leu Glu 180 185 190 Leu Ile
Arg Met Ile Lys Arg Gly Ile Asn Asp Arg Asn Phe Trp Arg 195 200 205
Gly Glu Asn Gly Arg Lys Thr Arg Ile Ala Tyr Glu Arg Met Cys Asn 210
215 220 Ile Leu Lys Gly Lys Phe Gln Thr Ala Ala Gln Lys Ala Met Met
Asp 225 230 235 240 Gln Val Arg Glu Ser Arg Asn Pro Gly Asn Ala Glu
Ile Glu Asp Leu 245 250 255 Thr Phe Leu Ala Arg Ser Ala Leu Ile Leu
Arg Gly Ser Val Ala His 260 265 270 Lys Ser Cys Leu Pro Ala Cys Val
Tyr Gly Pro Ala Val Ala Ser Gly 275 280 285 Tyr Asp Phe Glu Lys Glu
Gly Tyr Ser Leu Val Gly Val Asp Pro Phe 290 295 300 Lys Leu Leu Gln
Thr Ser Gln Val Tyr Ser Leu Ile Arg Pro Asn Glu 305 310 315 320 Asn
Pro Ala His Lys Ser Gln Leu Val Trp Met Ala Cys Asn Ser Ala 325 330
335 Ala Phe Glu Asp Leu Arg Val Ser Ser Phe Ile Arg Gly Thr Arg Val
340 345 350 Leu Pro Arg Gly Lys Leu Ser Thr Arg Gly Val Gln Ile Ala
Ser Asn 355 360 365 Glu Asn Met Asp Ala Ile Val Ser Ser Thr Leu Glu
Leu Arg Ser Arg 370 375 380 Tyr Trp Ala Ile Arg Thr Arg Ser Gly Gly
Asn Thr Asn Gln Gln Arg 385 390 395 400 Ala Ser Ala Gly Gln Ile Ser
Thr Gln Pro Thr Phe Ser Val Gln Arg 405 410 415 Asn Leu Pro Phe Asp
Lys Thr Thr Ile Met Ala Ala Phe Thr Gly Asn 420 425 430 Thr Glu Gly
Arg Thr Ser Asp Met Arg Ala Glu Ile Ile Lys Met Met 435 440 445 Glu
Ser Ala Arg Pro Glu Glu Val Ser Phe Gln Gly Arg Gly Val Phe 450 455
460 Glu Leu Ser Asp Glu Arg Ala Thr Asn Pro Ile Val Pro Ser Phe Asp
465 470 475 480 Met Ser Asn Glu Gly Ser Tyr Phe Phe Gly Asp Asn Ala
Glu Glu Tyr 485 490 495 Asp Asn 33252PRTInfluenza A virus 33Met Ser
Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser Ile Val Pro 1 5 10 15
Ser Gly Pro Leu Lys Ala Glu Ile Ala Gln Arg Leu Glu Asn Val Phe 20
25 30 Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys
Thr 35 40 45 Arg Pro Ile Leu Ser Pro Leu Thr Lys Gly Ile Leu Gly
Phe Val Phe 50 55 60 Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gln
Arg Arg Arg Phe Val 65 70 75 80 Gln Asn Ala Leu Asn Gly Asn Gly Asp
Pro Asn Asn Met Asp Arg Ala 85 90 95 Val Lys Leu Tyr Arg Lys Leu
Lys Arg Glu Ile Thr Phe His Gly Ala 100 105 110 Lys Glu Ile Ala Leu
Ser Tyr Ser Ala Gly Ala Leu Ala Ser Cys Met 115 120 125 Gly Leu Ile
Tyr Asn Arg Met Gly Ala Val Thr Thr Glu Ser Ala Phe 130 135 140 Gly
Leu Ile Cys Ala Thr Cys Glu Gln Ile Ala Asp Ser Gln His Lys 145 150
155 160 Ser His Arg Gln Met Val Thr Thr Thr Asn Pro Leu Ile Arg His
Glu 165 170 175 Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met
Glu Gln Met 180 185 190 Ala Gly Ser Ser Glu Gln Ala Ala Glu Ala Met
Glu Val Ala Ser Gln 195 200 205 Ala Arg Gln Met Val Gln Ala Met Arg
Ala Ile Gly Thr His Pro Ser 210 215 220 Ser Ser Thr Gly Leu Lys Asn
Asp Leu Leu Glu Asn Leu Gln Ala Tyr 225 230 235 240 Gln Lys Arg Met
Gly Val Gln Met Gln Arg Phe Lys 245 250 34470PRTInfluenza A virus
34Met Asn Pro Asn Gln Lys Ile Ile Thr Ile Gly Ser Ile Ser Ile Ala 1
5 10 15 Ile Gly Ile Ile Ser Leu Met Leu Gln Ile Gly Asn Ile Ile Ser
Ile 20 25 30 Trp Ala Ser His Ser Ile Gln Thr Gly Ser Gln Asn His
Thr Gly Val 35 40 45 Cys Asn Gln Arg Ile Ile Thr Tyr Glu Asn Ser
Thr Trp Val Asn His 50 55 60 Thr Tyr Val Asn Ile Asn Asn Thr Asn
Val Val Ala Gly Lys Asp Lys 65 70 75 80 Thr Ser Val Thr Leu Ala Gly
Asn Ser Ser Leu Cys Ser Ile Ser Gly 85 90 95 Trp Ala Ile Tyr Thr
Lys Asp Asn Ser Ile Arg Ile Gly Ser Lys Gly 100 105 110 Asp Val Phe
Val Ile Arg Glu Pro Phe Ile Ser Cys Ser His Leu Glu 115 120 125 Cys
Arg Thr Phe Phe Leu Thr Gln Gly Ala Leu Leu Asn Asp Lys His 130 135
140 Ser Asn Gly Thr Val Lys Asp Arg Ser Pro Tyr Arg Ala Leu Met Ser
145 150 155 160 Cys Pro Leu Gly Glu Ala Pro Ser Pro Tyr Asn Ser Lys
Phe Glu Ser 165 170 175 Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly
Met Gly Trp Leu Thr 180 185 190 Ile Gly Ile Ser Gly Pro Asp Asn Gly
Ala Val Ala Val Leu Lys Tyr 195 200 205 Asn Gly Ile Ile Thr Glu Thr
Ile Lys Ser Trp Lys Lys Arg Ile Leu 210
215 220 Arg Thr Gln Glu Ser Glu Cys Val Cys Val Asn Gly Ser Cys Phe
Thr 225 230 235 240 Ile Met Thr Asp Gly Pro Ser Asn Gly Ala Ala Ser
Tyr Lys Ile Phe 245 250 255 Lys Ile Glu Lys Gly Lys Val Thr Lys Ser
Ile Glu Leu Asn Ala Pro 260 265 270 Asn Phe His Tyr Glu Glu Cys Ser
Cys Tyr Pro Asp Thr Gly Thr Val 275 280 285 Met Cys Val Cys Arg Asp
Asn Trp His Gly Ser Asn Arg Pro Trp Val 290 295 300 Ser Phe Asn Gln
Asn Leu Asp Tyr Gln Ile Gly Tyr Ile Cys Ser Gly 305 310 315 320 Val
Phe Gly Asp Asn Pro Arg Pro Lys Asp Gly Glu Gly Ser Cys Asn 325 330
335 Pro Val Thr Val Asp Gly Ala Asp Gly Val Lys Gly Phe Ser Tyr Lys
340 345 350 Tyr Gly Asn Gly Val Trp Ile Gly Arg Thr Lys Ser Asn Arg
Leu Arg 355 360 365 Lys Gly Phe Glu Met Ile Trp Asp Pro Asn Gly Trp
Thr Asp Thr Asp 370 375 380 Ser Asp Phe Ser Val Lys Gln Asp Val Val
Ala Ile Thr Asp Trp Ser 385 390 395 400 Gly Tyr Ser Gly Ser Phe Val
Gln His Pro Glu Leu Thr Gly Leu Asp 405 410 415 Cys Ile Arg Pro Cys
Phe Trp Val Glu Leu Val Arg Gly Leu Pro Arg 420 425 430 Glu Asn Thr
Thr Ile Trp Thr Ser Gly Ser Ser Ile Ser Phe Cys Gly 435 440 445 Val
Asn Ser Asp Thr Ala Asn Trp Ser Trp Pro Asp Gly Ala Glu Leu 450 455
460 Pro Phe Thr Ile Asp Lys 465 470 35716PRTInfluenza A virus 35Met
Glu Asp Phe Val Arg Gln Cys Phe Asn Pro Met Ile Val Glu Leu 1 5 10
15 Ala Glu Lys Ala Met Lys Glu Tyr Gly Glu Asp Leu Lys Ile Glu Thr
20 25 30 Asn Lys Phe Ala Ala Ile Cys Thr His Leu Glu Val Cys Phe
Met Tyr 35 40 45 Ser Asp Phe His Phe Ile Asn Glu Gln Gly Glu Ser
Ile Val Val Glu 50 55 60 Leu Asp Asp Pro Asn Ala Leu Leu Lys His
Arg Phe Glu Ile Ile Glu 65 70 75 80 Gly Arg Asp Arg Thr Met Ala Trp
Thr Val Val Asn Ser Ile Cys Asn 85 90 95 Thr Thr Gly Ala Gly Lys
Pro Lys Phe Leu Pro Asp Leu Tyr Asp Tyr 100 105 110 Lys Glu Asn Arg
Phe Ile Glu Ile Gly Val Thr Arg Arg Glu Val His 115 120 125 Ile Tyr
Tyr Leu Glu Lys Ala Asn Lys Ile Lys Ser Glu Asn Thr His 130 135 140
Ile His Ile Phe Ser Phe Thr Gly Glu Glu Met Ala Thr Lys Ala Asp 145
150 155 160 Tyr Thr Leu Asp Glu Glu Ser Arg Ala Arg Ile Lys Thr Arg
Leu Phe 165 170 175 Thr Ile Arg Gln Glu Met Ala Asn Arg Gly Leu Trp
Asp Ser Phe Arg 180 185 190 Gln Ser Glu Arg Gly Glu Glu Thr Ile Glu
Glu Lys Phe Glu Ile Thr 195 200 205 Gly Thr Met Arg Arg Leu Ala Asp
Gln Ser Leu Pro Pro Asn Phe Ser 210 215 220 Cys Leu Glu Asn Phe Arg
Ala Tyr Val Asp Gly Phe Glu Pro Asn Gly 225 230 235 240 Cys Ile Glu
Gly Lys Leu Ser Gln Met Ser Lys Glu Val Asn Ala Gln 245 250 255 Ile
Glu Pro Phe Leu Lys Thr Thr Pro Arg Pro Ile Lys Leu Pro Asn 260 265
270 Gly Pro Pro Cys Tyr Gln Arg Ser Lys Phe Leu Leu Met Asp Ala Leu
275 280 285 Lys Leu Ser Ile Glu Asp Pro Ser His Glu Gly Glu Gly Ile
Pro Leu 290 295 300 Tyr Asp Ala Ile Lys Cys Met Lys Thr Phe Phe Gly
Trp Lys Glu Pro 305 310 315 320 Tyr Ile Val Lys Pro His Glu Lys Gly
Ile Asn Ser Asn Tyr Leu Leu 325 330 335 Ser Trp Lys Gln Val Leu Ser
Glu Leu Gln Asp Ile Glu Asn Glu Glu 340 345 350 Lys Ile Pro Arg Thr
Lys Asn Met Lys Lys Thr Ser Gln Leu Lys Trp 355 360 365 Ala Leu Gly
Glu Asn Met Ala Pro Glu Lys Val Asp Phe Glu Asn Cys 370 375 380 Arg
Asp Ile Ser Asp Leu Lys Gln Tyr Asp Ser Asp Glu Pro Glu Leu 385 390
395 400 Arg Ser Leu Ser Ser Trp Ile Gln Asn Glu Phe Asn Lys Ala Cys
Glu 405 410 415 Leu Thr Asp Ser Val Trp Ile Glu Leu Asp Glu Ile Gly
Glu Asp Val 420 425 430 Ala Pro Ile Glu His Ile Ala Ser Met Arg Arg
Asn Tyr Phe Thr Ala 435 440 445 Glu Val Ser His Cys Arg Ala Thr Glu
Tyr Ile Met Lys Gly Val Tyr 450 455 460 Ile Asn Thr Ala Leu Leu Asn
Ala Ser Cys Ala Ala Met Asp Asp Phe 465 470 475 480 Gln Leu Ile Pro
Met Ile Ser Lys Cys Arg Thr Lys Glu Gly Arg Arg 485 490 495 Lys Thr
Asn Leu Tyr Gly Phe Ile Ile Lys Gly Arg Ser His Leu Arg 500 505 510
Asn Asp Thr Asp Val Val Asn Phe Val Ser Met Glu Phe Ser Leu Thr 515
520 525 Asp Pro Arg Leu Glu Pro His Lys Trp Glu Lys Tyr Cys Val Leu
Glu 530 535 540 Ile Gly Asp Met Leu Leu Arg Ser Ala Ile Gly Gln Ile
Ser Arg Pro 545 550 555 560 Met Phe Leu Tyr Val Arg Thr Asn Gly Thr
Ser Lys Val Lys Met Lys 565 570 575 Trp Gly Met Glu Met Arg Arg Cys
Leu Leu Gln Ser Leu Gln Gln Ile 580 585 590 Glu Ser Met Ile Glu Ala
Glu Ser Ser Val Lys Glu Lys Asp Met Thr 595 600 605 Lys Glu Phe Phe
Glu Asn Lys Ser Glu Ala Trp Pro Ile Gly Glu Ser 610 615 620 Pro Lys
Gly Val Glu Glu Gly Ser Ile Gly Lys Val Cys Arg Thr Leu 625 630 635
640 Leu Ala Lys Ser Val Phe Asn Ser Leu Tyr Ala Ser Pro Gln Leu Glu
645 650 655 Gly Phe Ser Ala Glu Ser Arg Lys Leu Leu Leu Val Val Gln
Ala Leu 660 665 670 Arg Asp Asn Leu Glu Pro Gly Thr Phe Asp Leu Gly
Gly Leu Tyr Glu 675 680 685 Ala Ile Glu Glu Cys Leu Ile Asn Asp Pro
Trp Val Leu Leu Asn Ala 690 695 700 Ser Trp Phe Asn Ser Phe Leu Thr
His Ala Leu Lys 705 710 715 36757PRTInfluenza A virus 36Met Asp Val
Asn Pro Thr Leu Leu Phe Leu Lys Val Pro Ala Gln Asn 1 5 10 15 Ala
Ile Ser Thr Thr Phe Pro Tyr Thr Gly Asp Pro Pro Tyr Ser His 20 25
30 Gly Thr Gly Thr Gly Tyr Thr Met Asp Thr Val Asn Arg Thr His Gln
35 40 45 Tyr Ser Glu Lys Gly Lys Trp Thr Thr Asn Thr Glu Thr Gly
Ala Pro 50 55 60 Gln Leu Asn Pro Ile Asp Gly Pro Leu Pro Glu Asp
Asn Glu Pro Ser 65 70 75 80 Gly Tyr Ala Gln Thr Asp Cys Val Leu Glu
Ala Met Ala Phe Leu Glu 85 90 95 Glu Ser His Pro Gly Ile Phe Glu
Asn Ser Cys Leu Glu Thr Met Glu 100 105 110 Ala Val Gln Gln Thr Arg
Val Asp Arg Leu Thr Gln Gly Arg Gln Thr 115 120 125 Tyr Asp Trp Thr
Leu Asn Arg Asn Gln Pro Ala Ala Thr Ala Leu Ala 130 135 140 Asn Thr
Ile Glu Val Phe Arg Ser Asn Gly Leu Thr Ala Asn Glu Ser 145 150 155
160 Gly Arg Leu Ile Asp Phe Leu Lys Asp Val Met Glu Ser Met Asp Lys
165 170 175 Glu Glu Met Glu Ile Thr Thr His Phe Gln Arg Lys Arg Arg
Val Arg 180 185 190 Asp Asn Met Thr Lys Lys Met Val Thr Gln Arg Thr
Ile Gly Lys Lys 195 200 205 Lys Gln Arg Val Asn Lys Arg Gly Tyr Leu
Ile Arg Ala Leu Thr Leu 210 215 220 Asn Thr Met Thr Lys Asp Ala Glu
Arg Gly Lys Leu Lys Arg Arg Ala 225 230 235 240 Ile Ala Thr Pro Gly
Met Gln Ile Arg Gly Phe Val Tyr Phe Val Glu 245 250 255 Thr Leu Ala
Arg Ser Ile Cys Glu Lys Leu Glu Gln Ser Gly Leu Pro 260 265 270 Val
Gly Gly Asn Glu Lys Lys Ala Lys Leu Ala Asn Val Val Arg Lys 275 280
285 Met Met Thr Asn Ser Gln Asp Thr Glu Leu Ser Phe Thr Ile Thr Gly
290 295 300 Asp Asn Thr Lys Trp Asn Glu Asn Gln Asn Pro Arg Met Phe
Leu Ala 305 310 315 320 Met Ile Thr Tyr Ile Thr Lys Asn Gln Pro Glu
Trp Phe Arg Asn Ile 325 330 335 Leu Ser Ile Ala Pro Ile Met Phe Ser
Asn Lys Met Ala Arg Leu Gly 340 345 350 Lys Gly Tyr Met Phe Glu Ser
Lys Arg Met Lys Leu Arg Thr Gln Ile 355 360 365 Pro Ala Glu Met Leu
Ala Ser Ile Asp Leu Lys Tyr Phe Asn Glu Ser 370 375 380 Thr Arg Lys
Lys Ile Glu Lys Ile Arg Pro Leu Leu Ile Asp Gly Thr 385 390 395 400
Ala Ser Leu Ser Pro Gly Met Met Met Gly Met Phe Asn Met Leu Ser 405
410 415 Thr Val Leu Gly Val Ser Ile Leu Asn Leu Gly Gln Lys Lys Tyr
Thr 420 425 430 Lys Thr Thr Tyr Trp Trp Asp Gly Leu Gln Ser Ser Asp
Asp Phe Ala 435 440 445 Leu Ile Val Asn Ala Pro Asn His Glu Gly Ile
Gln Ala Gly Val Asn 450 455 460 Arg Phe Tyr Arg Thr Cys Lys Leu Val
Gly Ile Asn Met Ser Lys Lys 465 470 475 480 Lys Ser Tyr Ile Asn Lys
Thr Gly Thr Phe Glu Phe Thr Ser Phe Phe 485 490 495 Tyr Arg Tyr Gly
Phe Val Ala Asn Phe Ser Met Glu Leu Pro Ser Phe 500 505 510 Gly Val
Ser Gly Ile Asn Glu Ser Ala Asp Met Ser Ile Gly Val Thr 515 520 525
Val Ile Lys Asn Asn Met Ile Asn Asn Asp Leu Gly Pro Ala Thr Ala 530
535 540 Gln Met Ala Leu Gln Leu Phe Ile Lys Asp Tyr Arg Tyr Thr Tyr
Arg 545 550 555 560 Cys His Arg Gly Asp Thr Gln Ile Gln Thr Arg Arg
Ser Phe Glu Leu 565 570 575 Lys Lys Leu Trp Asp Gln Thr Gln Ser Arg
Ala Gly Leu Leu Val Ser 580 585 590 Asp Gly Gly Pro Asn Leu Tyr Asn
Ile Arg Asn Leu His Ile Pro Glu 595 600 605 Val Cys Leu Lys Trp Glu
Leu Met Asp Glu Asn Tyr Arg Gly Arg Leu 610 615 620 Cys Asn Pro Leu
Asn Pro Phe Val Ser His Lys Glu Ile Glu Ser Val 625 630 635 640 Asn
Asn Ala Val Val Met Pro Ala His Gly Pro Ala Lys Ser Met Glu 645 650
655 Tyr Asp Ala Val Ala Thr Thr His Ser Trp Ile Pro Lys Arg Asn Arg
660 665 670 Ser Ile Leu Asn Thr Ser Gln Arg Gly Ile Leu Glu Asp Glu
Gln Met 675 680 685 Tyr Gln Lys Cys Cys Asn Leu Phe Glu Lys Phe Phe
Pro Ser Ser Ser 690 695 700 Tyr Arg Arg Pro Ile Gly Ile Ser Ser Met
Val Glu Ala Met Val Ser 705 710 715 720 Arg Ala Arg Ile Asp Ala Arg
Ile Asp Phe Glu Ser Gly Arg Ile Lys 725 730 735 Lys Glu Glu Phe Ser
Glu Ile Met Lys Ile Cys Ser Thr Ile Glu Glu 740 745 750 Leu Arg Arg
Gln Arg 755 37759PRTInfluenza A virus 37Met Glu Arg Ile Lys Glu Leu
Arg Asn Leu Met Ser Gln Ser Arg Thr 1 5 10 15 Arg Glu Ile Leu Thr
Lys Thr Thr Val Asp His Met Ala Ile Ile Lys 20 25 30 Lys Tyr Thr
Ser Gly Arg Gln Glu Lys Asn Pro Ser Leu Arg Met Lys 35 40 45 Trp
Met Met Ala Met Lys Tyr Pro Ile Thr Ala Asp Lys Arg Ile Thr 50 55
60 Glu Met Val Pro Glu Arg Asn Glu Gln Gly Gln Thr Leu Trp Ser Lys
65 70 75 80 Met Ser Asp Ala Gly Ser Asp Arg Val Met Val Ser Pro Leu
Ala Val 85 90 95 Thr Trp Trp Asn Arg Asn Gly Pro Val Thr Ser Thr
Val His Tyr Pro 100 105 110 Lys Val Tyr Lys Thr Tyr Phe Asp Lys Val
Glu Arg Leu Lys His Gly 115 120 125 Thr Phe Gly Pro Val His Phe Arg
Asn Gln Val Lys Ile Arg Arg Arg 130 135 140 Val Asp Ile Asn Pro Gly
His Ala Asp Leu Ser Ala Lys Glu Ala Gln 145 150 155 160 Asp Val Ile
Met Glu Val Val Phe Pro Asn Glu Val Gly Ala Arg Ile 165 170 175 Leu
Thr Ser Glu Ser Gln Leu Thr Ile Thr Lys Glu Lys Lys Glu Glu 180 185
190 Leu Arg Asp Cys Lys Ile Ser Pro Leu Met Val Ala Tyr Met Leu Glu
195 200 205 Arg Glu Leu Val Arg Lys Thr Arg Phe Leu Pro Val Ala Gly
Gly Thr 210 215 220 Ser Ser Ile Tyr Ile Glu Val Leu His Leu Thr Gln
Gly Thr Cys Trp 225 230 235 240 Glu Gln Met Tyr Thr Pro Gly Gly Glu
Val Arg Asn Asp Asp Val Asp 245 250 255 Gln Ser Leu Ile Ile Ala Ala
Arg Asn Ile Val Arg Arg Ala Ala Val 260 265 270 Ser Ala Asp Pro Leu
Ala Ser Leu Leu Glu Met Cys His Ser Thr Gln 275 280 285 Ile Gly Gly
Thr Arg Met Val Asp Ile Leu Arg Gln Asn Pro Thr Glu 290 295 300 Glu
Gln Ala Val Asp Ile Cys Lys Ala Ala Met Gly Leu Arg Ile Ser 305 310
315 320 Ser Ser Phe Ser Phe Gly Gly Phe Thr Phe Lys Arg Thr Ser Gly
Ser 325 330 335 Ser Val Lys Lys Glu Glu Glu Val Leu Thr Gly Asn Leu
Gln Thr Leu 340 345 350 Lys Ile Arg Val His Glu Gly Tyr Glu Glu Phe
Thr Met Val Gly Lys 355 360 365 Arg Ala Thr Ala Ile Leu Arg Lys Ala
Thr Arg Arg Leu Val Gln Leu 370 375 380 Ile Val Ser Gly Arg Asp Glu
Gln Ser Ile Ala Glu Ala Ile Ile Val 385 390 395 400 Ala Met Val Phe
Ser Gln Glu Asp Cys Met Ile Lys Ala Val Arg Gly 405 410 415 Asp Leu
Asn Phe Val Asn Arg Ala Asn Gln Arg Leu Asn Pro Met His 420 425 430
Gln Leu Leu Arg His Phe Gln Lys Asp Ala Lys Val Leu Phe Gln Asn 435
440 445 Trp Gly Ile Glu His Ile Asp Ser Val Met Gly Met Val Gly Val
Leu 450 455 460 Pro Asp Met Thr Pro Ser Thr Glu Met Ser Met Arg Gly
Ile Arg Val 465 470 475 480 Ser Lys Met Gly Val Asp Glu Tyr Ser Ser
Thr Glu Arg Val Val Val 485 490 495 Ser Ile Asp Arg Phe Leu Arg Val
Arg Asp Gln Arg Gly Asn Val Leu 500 505 510 Leu Ser Pro Glu Glu Val
Ser Glu Thr Gln Gly Thr Glu Arg Leu Thr 515 520 525 Ile Thr Tyr Ser
Ser Ser Met Met Trp Glu Ile Asn Gly Pro Glu Ser 530 535 540 Val Leu
Val Asn Thr Tyr Gln Trp Ile Ile Arg Asn Trp Glu Ala Val 545 550 555
560 Lys Ile Gln Trp Ser Gln Asn Pro Ala Met Leu Tyr Asn Lys Met
Glu
565 570 575 Phe Glu Pro Phe Gln Ser Leu Val Pro Lys Ala Ile Arg Ser
Gln Tyr 580 585 590 Ser Gly Phe Val Arg Thr Leu Phe Gln Gln Met Arg
Asp Val Leu Gly 595 600 605 Thr Phe Asp Thr Thr Gln Ile Ile Lys Leu
Leu Pro Phe Ala Ala Ala 610 615 620 Pro Pro Lys Gln Ser Arg Met Gln
Phe Ser Ser Leu Thr Val Asn Val 625 630 635 640 Arg Gly Ser Gly Met
Arg Ile Leu Val Arg Gly Asn Ser Pro Val Phe 645 650 655 Asn Tyr Asn
Lys Thr Thr Lys Arg Leu Thr Ile Leu Gly Lys Asp Ala 660 665 670 Gly
Thr Leu Ile Glu Asp Pro Asp Glu Ser Thr Ser Gly Val Glu Ser 675 680
685 Ala Val Leu Arg Gly Phe Leu Ile Ile Gly Lys Glu Asp Arg Arg Tyr
690 695 700 Gly Pro Ala Leu Ser Ile Asn Glu Leu Ser Asn Leu Ala Lys
Gly Glu 705 710 715 720 Lys Ala Asn Val Leu Ile Gly Gln Gly Asp Val
Val Leu Val Met Lys 725 730 735 Arg Lys Arg Asp Ser Ser Ile Leu Thr
Asp Ser Gln Thr Ala Thr Lys 740 745 750 Arg Ile Arg Met Ala Ile Asn
755 38498PRTInfluenza A virus 38Met Ala Ser Gln Gly Thr Lys Arg Ser
Tyr Glu Gln Met Glu Thr Asp 1 5 10 15 Gly Asp Arg Gln Asn Ala Thr
Glu Ile Arg Ala Ser Val Gly Lys Met 20 25 30 Ile Asp Gly Ile Gly
Arg Phe Tyr Ile Gln Met Cys Thr Glu Leu Lys 35 40 45 Leu Ser Asp
Tyr Glu Gly Arg Leu Ile Gln Asn Ser Leu Thr Ile Glu 50 55 60 Lys
Met Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu 65 70
75 80 Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro
Ile 85 90 95 Tyr Arg Arg Val Asp Gly Lys Trp Met Arg Glu Leu Val
Leu Tyr Asp 100 105 110 Lys Glu Glu Ile Arg Arg Ile Trp Arg Gln Ala
Asn Asn Gly Glu Asp 115 120 125 Ala Thr Ala Gly Leu Thr His Ile Met
Ile Trp His Ser Asn Leu Asn 130 135 140 Asp Ala Thr Tyr Gln Arg Thr
Arg Ala Leu Val Arg Thr Gly Met Asp 145 150 155 160 Pro Arg Met Cys
Ser Leu Met Gln Gly Ser Thr Leu Pro Arg Arg Ser 165 170 175 Gly Ala
Ala Gly Ala Ala Val Lys Gly Ile Gly Thr Met Val Met Glu 180 185 190
Leu Ile Arg Met Val Lys Arg Gly Ile Asn Asp Arg Asn Phe Trp Arg 195
200 205 Gly Glu Asn Gly Arg Lys Thr Arg Ser Ala Tyr Glu Arg Met Cys
Asn 210 215 220 Ile Leu Lys Gly Lys Phe Gln Thr Ala Ala Gln Arg Ala
Met Val Asp 225 230 235 240 Gln Val Arg Glu Ser Arg Asn Pro Gly Asn
Ala Glu Ile Glu Asp Leu 245 250 255 Ile Phe Leu Ala Arg Ser Ala Leu
Ile Leu Arg Gly Ser Val Ala His 260 265 270 Lys Ser Cys Leu Pro Ala
Cys Val Tyr Gly Pro Ala Val Ser Ser Gly 275 280 285 Tyr Asn Phe Glu
Lys Glu Gly Tyr Ser Leu Val Gly Ile Asp Pro Phe 290 295 300 Lys Leu
Leu Gln Asn Ser Gln Val Tyr Ser Leu Ile Arg Pro Asn Glu 305 310 315
320 Asn Pro Ala His Lys Ser Gln Leu Val Trp Met Ala Cys His Ser Ala
325 330 335 Ala Phe Glu Asp Leu Arg Leu Leu Ser Phe Ile Arg Gly Thr
Lys Val 340 345 350 Ser Pro Arg Gly Lys Leu Ser Thr Arg Gly Val Gln
Ile Ala Ser Asn 355 360 365 Glu Asn Met Asp Asn Met Gly Ser Gly Thr
Leu Glu Leu Arg Ser Gly 370 375 380 Tyr Trp Ala Ile Arg Thr Arg Ser
Gly Gly Asn Thr Asn Gln Gln Arg 385 390 395 400 Ala Ser Ala Gly Gln
Thr Ser Val Gln Pro Thr Phe Ser Val Gln Arg 405 410 415 Asn Leu Pro
Phe Glu Lys Ser Thr Ile Met Ala Ala Phe Thr Gly Asn 420 425 430 Thr
Glu Gly Arg Thr Ser Asp Met Arg Ala Glu Ile Ile Arg Met Met 435 440
445 Glu Gly Ala Lys Pro Glu Glu Val Ser Phe Arg Gly Arg Gly Val Phe
450 455 460 Glu Leu Ser Asp Glu Lys Ala Thr Asn Pro Ile Val Pro Ser
Phe Asp 465 470 475 480 Met Ser Asn Glu Gly Ser Tyr Phe Phe Gly Asp
Asn Ala Glu Glu Tyr 485 490 495 Asp Asn 39252PRTInfluenza A virus
39Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser Ile Val Pro 1
5 10 15 Ser Gly Pro Leu Lys Ala Glu Ile Ala Gln Arg Leu Glu Asp Val
Phe 20 25 30 Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp
Leu Lys Thr 35 40 45 Arg Pro Ile Leu Ser Pro Leu Thr Lys Gly Ile
Leu Gly Phe Val Phe 50 55 60 Thr Leu Thr Val Pro Ser Glu Arg Gly
Leu Gln Arg Arg Arg Phe Val 65 70 75 80 Gln Asn Ala Leu Asn Gly Asn
Gly Asp Pro Asn Asn Met Asp Lys Ala 85 90 95 Val Lys Leu Tyr Arg
Lys Leu Lys Arg Glu Ile Thr Phe His Gly Ala 100 105 110 Lys Glu Ile
Ala Leu Ser Tyr Ser Ala Gly Ala Leu Ala Ser Cys Met 115 120 125 Gly
Leu Ile Tyr Asn Arg Met Gly Ala Val Thr Thr Glu Val Ala Phe 130 135
140 Gly Leu Val Cys Ala Thr Cys Glu Gln Ile Ala Asp Ser Gln His Arg
145 150 155 160 Ser His Arg Gln Met Val Ala Thr Thr Asn Pro Leu Ile
Arg His Glu 165 170 175 Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys
Ala Met Glu Gln Met 180 185 190 Ala Gly Ser Ser Glu Gln Ala Ala Glu
Ala Met Glu Ile Ala Ser Gln 195 200 205 Ala Arg Gln Met Val Gln Ala
Met Arg Ala Ile Gly Thr His Pro Ser 210 215 220 Ser Ser Thr Gly Leu
Arg Asp Asp Leu Leu Glu Asn Leu Gln Thr Tyr 225 230 235 240 Gln Lys
Arg Met Gly Val Gln Met Gln Arg Phe Lys 245 250 4097PRTInfluenza A
virus 40Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp
Gly 1 5 10 15 Cys Arg Cys Asn Asp Ser Ser Asp Pro Leu Val Val Ala
Ala Asn Ile 20 25 30 Ile Gly Ile Leu His Leu Ile Leu Trp Ile Leu
Asp Arg Leu Phe Phe 35 40 45 Lys Cys Val Tyr Arg Leu Phe Lys His
Gly Leu Lys Arg Gly Pro Ser 50 55 60 Thr Glu Gly Val Pro Glu Ser
Met Arg Glu Glu Tyr Arg Lys Glu Gln 65 70 75 80 Gln Asn Ala Val Asp
Ala Asp Asp Ser His Phe Val Ser Ile Glu Leu 85 90 95 Glu
41846DNAInfluenza A virus 41aatggattcc aacactgtgt caagtttcca
ggtagattgc tttctttggc atatccggaa 60acaagttgta gaccaagaac tgagtgatgc
cccattcctt gatcggcttc gccgagatca 120gaggtcccta aggggaagag
gcaatactct cggtctagac atcaaagcag ccacccatgt 180tggaaagcaa
attgtagaaa agattctgaa agaagaatct gatgaggcac ttaaaatgac
240catggtctcc acacctgctt cgcgatacat aactgacatg actattgagg
aattgtcaag 300aaactggttc atgctaatgc ccaagcagaa agtggaagga
cctctttgca tcagaatgga 360ccaggcaatc atggagaaaa acatcatgtt
gaaagcgaat ttcagtgtga tttctgaccg 420actagagacc atagtattac
taagggcttt caccgaagag ggagcaattg ttggcgaaat 480ctcaccattg
ccttcttttc caggacatac tattgaggat gtcaaaaatg caattggggt
540cctcatcgga ggacttgaat ggaatgataa cacagttcga gtctctaaaa
atctacagag 600attcgcttgg agaagcagta atgagaatgg gggacctcca
cttactccaa aacagaaacg 660gaaaatggcg agaacagcta ggtcaaaagt
ttgaagagat aagatggctg attgaagaag 720tgagacacag actaaaaaca
actgaaaata gctttgaaca aataacattc atgcaagcat 780tacaactgct
gtttgaagtg gaacaggaga taagaacttt ctcatttcag cttatttaat 840gataaa
84642566PRTInfluenza A virus 42Met Lys Thr Ile Ile Ala Leu Ser Tyr
Ile Leu Cys Leu Val Phe Ala 1 5 10 15 Gln Lys Leu Pro Gly Asn Asp
Asn Ser Thr Ala Thr Leu Cys Leu Gly 20 25 30 His His Ala Val Pro
Asn Gly Thr Ile Val Lys Thr Ile Thr Asn Asp 35 40 45 Gln Ile Glu
Val Thr Asn Ala Thr Glu Leu Val Gln Ser Ser Ser Thr 50 55 60 Gly
Gly Ile Cys Asp Ser Pro His Gln Ile Leu Asp Gly Glu Asn Cys 65 70
75 80 Thr Leu Ile Asp Ala Leu Leu Gly Asp Pro Gln Cys Asp Gly Phe
Gln 85 90 95 Asn Lys Lys Trp Asp Leu Phe Val Glu Arg Ser Lys Ala
Tyr Ser Asn 100 105 110 Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser
Leu Arg Ser Leu Val 115 120 125 Ala Ser Ser Gly Thr Leu Glu Phe Asn
Asp Glu Ser Phe Asn Trp Thr 130 135 140 Gly Val Thr Gln Asn Gly Thr
Ser Ser Ser Cys Lys Arg Arg Ser Asn 145 150 155 160 Asn Ser Phe Phe
Ser Arg Leu Asn Trp Leu Thr His Leu Lys Phe Lys 165 170 175 Tyr Pro
Ala Leu Asn Val Thr Met Pro Asn Asn Glu Lys Phe Asp Lys 180 185 190
Leu Tyr Ile Trp Gly Val His His Pro Val Thr Asp Asn Asp Gln Ile 195
200 205 Phe Leu Tyr Ala Gln Ala Ser Gly Arg Ile Thr Val Ser Thr Lys
Arg 210 215 220 Ser Gln Gln Thr Val Ile Pro Asn Ile Gly Ser Arg Pro
Arg Ile Arg 225 230 235 240 Asn Ile Pro Ser Arg Ile Ser Ile Tyr Trp
Thr Ile Val Lys Pro Gly 245 250 255 Asp Ile Leu Leu Ile Asn Ser Thr
Gly Asn Leu Ile Ala Pro Arg Gly 260 265 270 Tyr Phe Lys Ile Arg Ser
Gly Lys Ser Ser Ile Met Arg Ser Asp Ala 275 280 285 Pro Ile Gly Lys
Cys Asn Ser Glu Cys Ile Thr Pro Asn Gly Ser Ile 290 295 300 Pro Asn
Asp Lys Pro Phe Gln Asn Val Asn Arg Ile Thr Tyr Gly Ala 305 310 315
320 Cys Pro Arg Tyr Val Lys Gln Asn Thr Leu Lys Leu Ala Thr Gly Met
325 330 335 Arg Asn Val Pro Glu Lys Gln Thr Arg Gly Ile Phe Gly Ala
Ile Ala 340 345 350 Gly Phe Ile Glu Asn Gly Trp Glu Gly Met Val Asp
Gly Trp Tyr Gly 355 360 365 Phe Arg His Gln Asn Ser Glu Gly Ile Gly
Gln Ala Ala Asp Leu Lys 370 375 380 Ser Thr Gln Ala Ala Ile Asn Gln
Ile Asn Gly Lys Leu Asn Arg Leu 385 390 395 400 Ile Gly Lys Thr Asn
Glu Lys Phe His Gln Ile Glu Lys Glu Phe Ser 405 410 415 Glu Val Glu
Gly Arg Ile Gln Asp Leu Glu Lys Tyr Val Glu Asp Thr 420 425 430 Lys
Ile Asp Leu Trp Ser Tyr Asn Ala Glu Leu Leu Val Ala Leu Glu 435 440
445 Asn Gln His Thr Ile Asp Leu Thr Asp Ser Glu Met Asn Lys Leu Phe
450 455 460 Glu Arg Thr Lys Lys Gln Leu Arg Glu Asn Ala Glu Asp Met
Gly Asn 465 470 475 480 Gly Cys Phe Lys Ile Tyr His Lys Cys Asp Asn
Ala Cys Ile Gly Ser 485 490 495 Ile Arg Asn Gly Thr Tyr Asp His Asp
Val Tyr Arg Asp Glu Ala Leu 500 505 510 Asn Asn Arg Phe Gln Ile Lys
Gly Val Glu Leu Lys Ser Gly Tyr Lys 515 520 525 Asp Trp Ile Leu Trp
Ile Ser Phe Ala Ile Ser Cys Phe Leu Leu Cys 530 535 540 Val Ala Leu
Leu Gly Phe Ile Met Trp Ala Cys Gln Lys Gly Asn Ile 545 550 555 560
Arg Cys Asn Ile Cys Ile 565 43469PRTInfluenza A virus 43Met Asn Pro
Asn Gln Lys Ile Ile Thr Ile Gly Ser Val Ser Leu Thr 1 5 10 15 Ile
Ser Thr Ile Cys Phe Phe Met Gln Ile Ala Ile Leu Ile Thr Thr 20 25
30 Val Thr Leu His Phe Lys Gln Tyr Glu Phe Asn Ser Pro Pro Asn Asn
35 40 45 Gln Val Met Leu Cys Glu Pro Thr Ile Ile Glu Arg Asn Ile
Thr Glu 50 55 60 Ile Val Tyr Leu Thr Asn Thr Thr Ile Glu Lys Glu
Ile Cys Pro Lys 65 70 75 80 Leu Ala Glu Tyr Arg Asn Trp Ser Lys Pro
Gln Cys Asn Ile Thr Gly 85 90 95 Phe Ala Pro Phe Ser Lys Asp Asn
Ser Ile Arg Leu Ser Ala Gly Gly 100 105 110 Asp Ile Trp Val Thr Arg
Glu Pro Tyr Val Ser Cys Asp Pro Asp Lys 115 120 125 Cys Tyr Gln Phe
Ala Leu Gly Gln Gly Thr Thr Leu Asn Asn Val His 130 135 140 Ser Asn
Asp Thr Val His Asp Arg Thr Pro Tyr Arg Thr Leu Leu Met 145 150 155
160 Asn Glu Leu Gly Val Pro Phe His Leu Gly Thr Lys Gln Val Cys Ile
165 170 175 Ala Trp Ser Ser Ser Ser Cys His Asp Gly Lys Ala Trp Leu
His Val 180 185 190 Cys Val Thr Gly Asp Asp Lys Asn Ala Thr Ala Ser
Phe Ile Tyr Asn 195 200 205 Gly Arg Leu Val Asp Ser Ile Val Ser Trp
Ser Lys Glu Ile Leu Arg 210 215 220 Thr Gln Glu Ser Glu Cys Val Cys
Ile Asn Gly Thr Cys Thr Val Val 225 230 235 240 Met Thr Asp Gly Ser
Ala Ser Gly Lys Ala Asp Thr Lys Ile Leu Phe 245 250 255 Ile Glu Glu
Gly Lys Ile Val His Thr Ser Thr Leu Ser Gly Ser Ala 260 265 270 Gln
His Val Glu Glu Cys Ser Cys Tyr Pro Arg Tyr Leu Gly Val Arg 275 280
285 Cys Val Cys Arg Asp Asn Trp Lys Gly Ser Asn Arg Pro Ile Val Asp
290 295 300 Ile Asn Ile Lys Asp Tyr Ser Ile Val Ser Ser Tyr Val Cys
Ser Gly 305 310 315 320 Leu Val Gly Asp Thr Pro Arg Lys Asn Asp Ser
Ser Ser Ser Ser His 325 330 335 Cys Leu Asp Pro Asn Asn Glu Glu Gly
Gly His Gly Val Lys Gly Trp 340 345 350 Ala Phe Asp Asp Gly Asn Asp
Val Trp Met Gly Arg Thr Ile Ser Glu 355 360 365 Lys Leu Arg Ser Gly
Tyr Glu Thr Phe Lys Val Ile Glu Gly Trp Ser 370 375 380 Asn Pro Asn
Ser Lys Leu Gln Ile Asn Arg Gln Val Ile Val Asp Arg 385 390 395 400
Gly Asn Arg Ser Gly Tyr Ser Gly Ile Phe Ser Val Glu Gly Lys Ser 405
410 415 Cys Ile Asn Arg Cys Phe Tyr Val Glu Leu Ile Arg Gly Arg Lys
Glu 420 425 430 Glu Thr Glu Val Leu Trp Thr Ser Asn Ser Ile Val Val
Phe Cys Gly 435 440 445 Thr Ser Gly Thr Tyr Gly Thr Gly Ser Trp Pro
Asp Gly Ala Asp Ile 450 455 460 Asn Leu Met Pro Ile 465
44716PRTInfluenza A virus 44Met Glu Asp Phe Val Arg Gln Cys Phe Asn
Pro Met Ile Val Glu Leu 1 5 10 15 Ala Glu Lys Ala Met Lys Glu Tyr
Gly Glu Asp Pro Lys Ile Glu Thr 20 25 30 Asn Lys Phe Ala Ala Ile
Cys Thr His Leu Glu Val Cys Phe Met Tyr 35 40 45 Ser Asp Phe His
Phe Ile Asp Glu Arg Gly Glu Ser Ile Ile Val Glu 50 55 60 Ser Gly
Asp Pro Asn Ala Leu Leu Lys His Arg Phe Glu Ile Ile Glu 65
70 75 80 Gly Arg Asp Arg Ile Met Ala Trp Thr Val Ile Asn Ser Ile
Cys Asn 85 90 95 Thr Thr Gly Val Glu Lys Pro Lys Phe Leu Pro Asp
Leu Tyr Asp Tyr 100 105 110 Lys Glu Asn Arg Phe Ile Glu Ile Gly Val
Thr Arg Arg Glu Val His 115 120 125 Ile Tyr Tyr Leu Glu Lys Ala Asn
Lys Ile Lys Ser Glu Lys Thr His 130 135 140 Ile His Ile Phe Ser Phe
Thr Gly Glu Glu Met Ala Thr Lys Ala Asp 145 150 155 160 Tyr Thr Leu
Asp Glu Glu Ser Arg Ala Arg Ile Lys Thr Arg Leu Phe 165 170 175 Thr
Ile Arg Gln Glu Met Ala Ser Lys Ser Leu Trp Asp Ser Phe Arg 180 185
190 Gln Ser Glu Arg Gly Glu Glu Thr Ile Glu Glu Lys Phe Glu Ile Thr
195 200 205 Gly Thr Met Arg Lys Leu Ala Asp Gln Ser Leu Pro Pro Asn
Phe Pro 210 215 220 Ser Leu Glu Asn Phe Arg Ala Tyr Val Asp Gly Phe
Glu Pro Asn Gly 225 230 235 240 Cys Ile Glu Gly Lys Leu Ser Gln Met
Ser Lys Glu Val Asn Ala Lys 245 250 255 Ile Glu Pro Phe Leu Arg Thr
Thr Pro Arg Pro Leu Arg Leu Pro Asp 260 265 270 Gly Pro Leu Cys His
Gln Arg Ser Lys Phe Leu Leu Met Asp Ala Leu 275 280 285 Lys Leu Ser
Ile Glu Asp Pro Ser His Glu Gly Glu Gly Ile Pro Leu 290 295 300 Tyr
Asp Ala Ile Lys Cys Met Lys Thr Phe Phe Gly Trp Lys Glu Pro 305 310
315 320 Asn Ile Val Lys Pro His Glu Lys Gly Ile Asn Pro Asn Tyr Leu
Met 325 330 335 Ala Trp Lys Gln Val Leu Ala Glu Leu Gln Asp Ile Glu
Asn Glu Glu 340 345 350 Lys Ile Pro Arg Thr Lys Asn Met Lys Arg Thr
Ser Gln Leu Lys Trp 355 360 365 Ala Leu Gly Glu Asn Met Ala Pro Glu
Lys Val Asp Phe Asp Asp Cys 370 375 380 Lys Asp Val Gly Asp Leu Lys
Gln Tyr Asp Ser Asp Glu Pro Glu Pro 385 390 395 400 Arg Ser Leu Ala
Ser Trp Val Gln Asn Glu Phe Asn Lys Ala Cys Glu 405 410 415 Leu Thr
Asp Ser Ser Trp Ile Glu Leu Asp Glu Ile Gly Glu Asp Val 420 425 430
Ala Pro Ile Glu His Ile Ala Ser Met Arg Arg Asn Tyr Phe Thr Ala 435
440 445 Glu Val Ser His Cys Arg Ala Thr Glu Tyr Ile Met Lys Gly Val
Tyr 450 455 460 Ile Asn Thr Ala Leu Leu Asn Ala Ser Cys Ala Ala Met
Asp Asp Phe 465 470 475 480 Gln Leu Ile Pro Met Ile Ser Lys Cys Arg
Thr Lys Glu Gly Arg Arg 485 490 495 Lys Thr Asn Leu Tyr Gly Phe Ile
Ile Lys Gly Arg Ser His Leu Arg 500 505 510 Asn Asp Thr Asp Val Val
Asn Phe Val Ser Met Glu Phe Ser Leu Thr 515 520 525 Asp Pro Arg Leu
Glu Pro His Lys Trp Glu Lys Tyr Cys Val Leu Glu 530 535 540 Ile Gly
Asp Met Leu Leu Arg Thr Ala Ile Gly Gln Val Ser Arg Pro 545 550 555
560 Met Phe Leu Tyr Val Arg Thr Asn Gly Thr Ser Lys Ile Lys Met Lys
565 570 575 Trp Gly Met Glu Met Arg Arg Cys Leu Leu Gln Ser Leu Gln
Gln Ile 580 585 590 Glu Ser Met Ile Glu Ala Glu Ser Ser Val Lys Glu
Lys Asp Met Thr 595 600 605 Lys Glu Phe Phe Glu Asn Lys Ser Glu Thr
Trp Pro Ile Gly Glu Ser 610 615 620 Pro Arg Gly Val Glu Glu Gly Ser
Ile Gly Lys Val Cys Arg Thr Leu 625 630 635 640 Leu Ala Lys Ser Val
Phe Asn Ser Leu Tyr Ala Ser Pro Gln Leu Glu 645 650 655 Gly Phe Ser
Ala Glu Ser Arg Lys Leu Leu Leu Ile Val Gln Ala Leu 660 665 670 Arg
Asp Asn Leu Glu Pro Gly Thr Phe Asp Leu Gly Gly Leu Tyr Glu 675 680
685 Ala Ile Glu Glu Cys Leu Ile Asn Asp Pro Trp Val Leu Leu Asn Ala
690 695 700 Ser Trp Phe Asn Ser Phe Leu Thr His Ala Leu Lys 705 710
715 45252PRTInfluenza A virus 45Met Ser Leu Leu Thr Glu Val Glu Thr
Tyr Val Leu Ser Ile Val Pro 1 5 10 15 Ser Gly Pro Leu Lys Ala Glu
Ile Ala Gln Arg Leu Glu Asn Val Phe 20 25 30 Ala Gly Lys Asn Thr
Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr 35 40 45 Arg Pro Ile
Leu Ser Pro Leu Thr Lys Gly Ile Leu Gly Phe Val Phe 50 55 60 Thr
Leu Thr Val Pro Ser Glu Arg Gly Leu Gln Arg Arg Arg Phe Val 65 70
75 80 Gln Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Lys
Ala 85 90 95 Val Lys Leu Tyr Arg Lys Leu Lys Arg Glu Ile Thr Phe
His Gly Ala 100 105 110 Lys Glu Ile Ala Leu Ser Tyr Ser Ala Gly Ala
Leu Ala Ser Cys Met 115 120 125 Gly Leu Ile Tyr Asn Arg Met Gly Ala
Val Thr Thr Glu Ser Ala Phe 130 135 140 Gly Leu Ile Cys Ala Thr Cys
Glu Gln Ile Ala Asp Ser Gln His Lys 145 150 155 160 Ser His Arg Gln
Met Val Thr Thr Thr Asn Pro Leu Ile Arg His Glu 165 170 175 Asn Arg
Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gln Met 180 185 190
Ala Gly Ser Ser Glu Gln Ala Ala Glu Ala Met Glu Val Ala Ser Gln 195
200 205 Ala Arg Gln Met Val Gln Ala Met Arg Ala Ile Gly Thr His Pro
Ser 210 215 220 Ser Ser Thr Gly Leu Lys Asn Asp Leu Leu Glu Asn Leu
Gln Ala Tyr 225 230 235 240 Gln Lys Arg Met Gly Val Gln Met Gln Arg
Phe Lys 245 250 46758PRTInfluenza A virus 46Met Asp Val Asn Pro Thr
Leu Leu Phe Leu Lys Ile Pro Ala Gln Asn 1 5 10 15 Ala Ile Ser Thr
Thr Phe Pro Tyr Thr Gly Asp Pro Pro Tyr Ser His 20 25 30 Gly Thr
Gly Thr Gly Tyr Thr Met Asp Thr Val Asn Arg Thr His Gln 35 40 45
Tyr Ser Glu Lys Gly Lys Trp Thr Thr Asn Thr Glu Thr Gly Ala Pro 50
55 60 Gln Leu Asn Pro Ile Asp Gly Pro Leu Pro Glu Asp Asn Glu Pro
Ser 65 70 75 80 Gly Tyr Ala Gln Thr Asp Cys Val Leu Glu Ala Met Ala
Phe Leu Glu 85 90 95 Glu Ser His Pro Gly Ile Phe Glu Asn Ser Cys
Leu Glu Thr Met Glu 100 105 110 Val Val Gln Gln Thr Arg Val Asp Arg
Leu Thr Gln Gly Arg Gln Thr 115 120 125 Tyr Asp Trp Thr Leu Asn Arg
Asn Gln Pro Ala Ala Thr Ala Leu Ala 130 135 140 Asn Thr Ile Glu Val
Phe Arg Ser Asn Gly Leu Thr Ala Asn Glu Ser 145 150 155 160 Gly Arg
Leu Ile Asp Phe Leu Lys Asp Val Met Glu Ser Met Asp Lys 165 170 175
Glu Glu Ile Glu Ile Thr Thr His Phe Gln Arg Lys Arg Arg Val Arg 180
185 190 Asp Asn Met Thr Lys Lys Met Val Thr Gln Arg Thr Ile Gly Lys
Lys 195 200 205 Lys Gln Arg Val Asn Lys Arg Ser Tyr Leu Ile Arg Ala
Leu Thr Leu 210 215 220 Asn Thr Met Thr Lys Asp Ala Glu Arg Gly Lys
Leu Lys Arg Arg Ala 225 230 235 240 Ile Ala Thr Pro Gly Met Gln Ile
Arg Gly Phe Val Tyr Phe Val Glu 245 250 255 Thr Leu Ala Arg Ser Ile
Cys Glu Lys Leu Glu Gln Ser Gly Leu Pro 260 265 270 Val Gly Gly Asn
Glu Lys Lys Ala Lys Leu Ala Asn Val Val Arg Lys 275 280 285 Met Met
Thr Asn Ser Gln Asp Thr Glu Leu Ser Phe Thr Ile Thr Gly 290 295 300
Asp Asn Thr Lys Trp Asn Glu Asn Gln Asn Pro Arg Met Phe Leu Ala 305
310 315 320 Met Ile Thr Tyr Ile Thr Lys Asn Gln Pro Glu Trp Phe Arg
Asn Ile 325 330 335 Leu Ser Ile Ala Pro Ile Met Phe Ser Asn Lys Met
Ala Arg Leu Gly 340 345 350 Lys Gly Tyr Met Phe Glu Ser Lys Arg Met
Lys Leu Arg Thr Gln Ile 355 360 365 Pro Ala Glu Met Leu Ala Ser Ile
Asp Leu Lys Tyr Phe Asn Glu Ser 370 375 380 Thr Arg Lys Lys Ile Glu
Lys Ile Arg Pro Leu Leu Ile Asp Gly Thr 385 390 395 400 Ala Ser Leu
Ser Pro Gly Met Met Met Gly Met Phe Asn Met Leu Ser 405 410 415 Thr
Val Leu Gly Val Ser Ile Leu Asn Leu Gly Gln Lys Lys Tyr Thr 420 425
430 Lys Thr Thr Tyr Trp Trp Asp Gly Leu Gln Ser Ser Asp Asp Phe Ala
435 440 445 Leu Ile Val Asn Ala Pro Asn His Glu Gly Ile Gln Ala Gly
Val Asp 450 455 460 Arg Phe Tyr Arg Thr Cys Lys Leu Val Gly Ile Asn
Met Ser Lys Lys 465 470 475 480 Lys Ser Tyr Ile Asn Arg Thr Gly Thr
Phe Glu Phe Thr Ser Phe Phe 485 490 495 Tyr Arg Tyr Gly Phe Val Ala
Asn Phe Ser Met Glu Leu Pro Ser Phe 500 505 510 Gly Val Ser Gly Ile
Asn Glu Ser Ala Asp Met Ser Ile Gly Val Thr 515 520 525 Val Ile Lys
Asn Asn Met Ile Asn Asn Asp Leu Gly Pro Ala Thr Ala 530 535 540 Gln
Met Ala Leu Gln Leu Phe Ile Lys Asp Tyr Arg Tyr Thr Tyr Arg 545 550
555 560 Cys His Arg Gly Asp Thr Gln Ile Gln Thr Arg Arg Ser Phe Glu
Leu 565 570 575 Lys Lys Leu Trp Glu Gln Thr Arg Ser Lys Ala Gly Leu
Leu Val Ser 580 585 590 Asp Gly Gly Pro Asn Leu Tyr Asn Ile Arg Asn
Leu His Ile Pro Glu 595 600 605 Val Cys Leu Lys Trp Glu Leu Met Asp
Glu Asp Tyr Gln Gly Arg Leu 610 615 620 Cys Asn Pro Leu Asn Pro Phe
Val Ser His Lys Glu Ile Glu Ser Val 625 630 635 640 Asn Asn Ala Val
Val Met Pro Ala His Gly Pro Ala Lys Ser Met Glu 645 650 655 Tyr Asp
Ala Val Ala Thr Thr His Ser Trp Ile Pro Lys Arg Asn Arg 660 665 670
Ser Ile Leu Asn Thr Ser Gln Arg Gly Ile Leu Glu Asp Glu Gln Met 675
680 685 Tyr Gln Lys Cys Cys Asn Leu Phe Glu Lys Phe Phe Pro Ser Ser
Ser 690 695 700 Tyr Arg Arg Pro Val Gly Ile Ser Ser Met Val Glu Ala
Met Val Ser 705 710 715 720 Arg Ala Arg Ile Asp Ala Arg Ile Asp Phe
Glu Ser Gly Arg Ile Lys 725 730 735 Lys Glu Glu Phe Ser Glu Ile Met
Lys Ile Cys Ser Thr Ile Glu Glu 740 745 750 Leu Arg Arg Gln Lys Gln
755 47716PRTInfluenza A virus 47Met Glu Asp Phe Val Arg Gln Cys Phe
Asn Pro Met Ile Val Glu Leu 1 5 10 15 Ala Glu Lys Thr Met Lys Glu
Tyr Gly Glu Asp Leu Lys Ile Glu Thr 20 25 30 Asn Lys Phe Ala Ala
Ile Cys Thr His Leu Glu Val Cys Phe Met Tyr 35 40 45 Ser Asp Phe
His Phe Ile Asn Glu Gln Gly Glu Ser Ile Ile Val Glu 50 55 60 Leu
Gly Asp Pro Asn Ala Leu Leu Lys His Arg Phe Glu Ile Ile Glu 65 70
75 80 Gly Arg Asp Arg Thr Met Ala Trp Thr Val Val Asn Ser Ile Cys
Asn 85 90 95 Thr Thr Gly Ala Glu Lys Pro Lys Phe Leu Pro Asp Leu
Tyr Asp Tyr 100 105 110 Lys Glu Asn Arg Phe Ile Glu Ile Gly Val Thr
Arg Arg Glu Val His 115 120 125 Ile Tyr Tyr Leu Glu Lys Ala Asn Lys
Ile Lys Ser Glu Lys Thr His 130 135 140 Ile His Ile Phe Ser Phe Thr
Gly Glu Glu Met Ala Thr Lys Ala Asp 145 150 155 160 Tyr Thr Leu Asp
Glu Glu Ser Arg Ala Arg Ile Lys Thr Arg Leu Phe 165 170 175 Thr Ile
Arg Gln Glu Met Ala Ser Arg Gly Leu Trp Asp Ser Phe Arg 180 185 190
Gln Ser Glu Arg Gly Glu Glu Thr Ile Glu Glu Arg Phe Glu Ile Thr 195
200 205 Gly Thr Met Arg Lys Leu Ala Asp Gln Ser Leu Pro Pro Asn Phe
Ser 210 215 220 Ser Leu Glu Asn Phe Arg Ala Tyr Val Asp Gly Phe Glu
Pro Asn Gly 225 230 235 240 Tyr Ile Glu Gly Lys Leu Ser Gln Met Ser
Lys Glu Val Asn Ala Arg 245 250 255 Ile Glu Pro Phe Leu Lys Thr Thr
Pro Arg Pro Leu Arg Leu Pro Asn 260 265 270 Gly Pro Pro Cys Ser Gln
Arg Ser Lys Phe Leu Leu Met Asp Ala Leu 275 280 285 Lys Leu Ser Ile
Glu Asp Pro Ser His Glu Gly Glu Gly Ile Pro Leu 290 295 300 Tyr Asp
Ala Ile Lys Cys Met Arg Thr Phe Phe Gly Trp Lys Glu Pro 305 310 315
320 Asn Val Val Lys Pro His Glu Lys Gly Ile Asn Pro Asn Tyr Leu Leu
325 330 335 Ser Trp Lys Gln Val Leu Ala Glu Leu Gln Asp Ile Glu Asn
Glu Glu 340 345 350 Lys Ile Pro Lys Thr Lys Asn Met Lys Lys Thr Ser
Gln Leu Lys Trp 355 360 365 Ala Leu Gly Glu Asn Met Ala Pro Glu Lys
Val Asp Phe Asp Asp Cys 370 375 380 Lys Asp Val Gly Asp Leu Lys Gln
Tyr Asp Ser Asp Glu Pro Glu Leu 385 390 395 400 Arg Ser Leu Ala Ser
Trp Ile Gln Asn Glu Phe Asn Lys Ala Cys Glu 405 410 415 Leu Thr Asp
Ser Ser Trp Ile Glu Leu Asp Glu Ile Gly Glu Asp Val 420 425 430 Ala
Pro Ile Glu His Ile Ala Ser Met Arg Arg Asn Tyr Phe Thr Ser 435 440
445 Glu Val Ser His Cys Arg Ala Thr Glu Tyr Ile Met Lys Gly Val Tyr
450 455 460 Ile Asn Thr Ala Leu Leu Asn Ala Ser Cys Ala Ala Met Asp
Asp Phe 465 470 475 480 Gln Leu Ile Pro Met Ile Ser Lys Cys Arg Thr
Lys Glu Gly Arg Arg 485 490 495 Lys Thr Asn Leu Tyr Gly Phe Ile Ile
Lys Gly Arg Ser His Leu Arg 500 505 510 Asn Asp Thr Asp Val Val Asn
Phe Val Ser Met Glu Phe Ser Leu Thr 515 520 525 Asp Pro Arg Leu Glu
Pro His Lys Trp Glu Lys Tyr Cys Val Leu Glu 530 535 540 Ile Gly Asp
Met Leu Ile Arg Ser Ala Ile Gly Gln Val Ser Arg Pro 545 550 555 560
Met Phe Leu Tyr Val Arg Thr Asn Gly Thr Ser Lys Ile Lys Met Lys 565
570 575 Trp Gly Met Glu Met Arg Arg Cys Leu Leu Gln Ser Leu Gln Gln
Ile 580 585 590 Glu Ser Met Ile Glu Ala Glu Ser Ser Val Lys Glu Lys
Asp Met Thr 595 600 605 Lys Glu Phe Phe Glu Asn Lys Ser Glu Thr Trp
Pro Ile Gly Glu Ser 610 615 620 Pro Lys Gly Val Glu Glu Ser Ser Ile
Gly Lys Val Cys Arg Thr Leu 625 630 635 640 Leu Ala Lys Ser Val Phe
Asn Ser Leu
Tyr Ala Ser Pro Gln Leu Glu 645 650 655 Gly Phe Ser Ala Glu Ser Arg
Lys Leu Leu Leu Ile Val Gln Ala Leu 660 665 670 Arg Asp Asn Leu Glu
Pro Gly Thr Phe Asp Leu Gly Gly Leu Tyr Glu 675 680 685 Ala Ile Glu
Glu Cys Leu Ile Asn Asp Pro Trp Val Leu Leu Asn Ala 690 695 700 Ser
Trp Phe Asn Ser Phe Leu Thr His Ala Leu Ser 705 710 715
48326PRTInfluenza A virus 48Met Ala Ser Gln Gly Thr Lys Arg Ser Tyr
Glu Gln Met Glu Thr Asp 1 5 10 15 Gly Glu Arg Gln Asn Ala Thr Glu
Ile Arg Ala Ser Val Gly Lys Met 20 25 30 Ile Gly Gly Ile Gly Arg
Phe Tyr Ile Gln Met Cys Thr Glu Leu Lys 35 40 45 Leu Ser Asp Tyr
Glu Gly Arg Leu Ile Gln Asn Ser Leu Thr Ile Glu 50 55 60 Arg Met
Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu 65 70 75 80
Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro Ile 85
90 95 Tyr Arg Arg Val Asn Gly Lys Trp Met Arg Glu Leu Ile Leu Tyr
Asp 100 105 110 Lys Glu Glu Ile Arg Arg Ile Trp Arg Gln Ala Asn Asn
Gly Asp Asp 115 120 125 Ala Thr Ala Gly Leu Thr His Met Met Ile Trp
His Ser Asn Leu Asn 130 135 140 Asp Ala Thr Tyr Gln Arg Thr Arg Ala
Leu Val Arg Thr Gly Met Asp 145 150 155 160 Pro Arg Met Cys Ser Leu
Met Gln Gly Ser Thr Leu Pro Arg Arg Ser 165 170 175 Gly Ala Ala Gly
Ala Ala Val Lys Gly Val Gly Thr Met Val Met Glu 180 185 190 Leu Val
Arg Met Ile Lys Arg Gly Ile Asn Asp Arg Asn Phe Trp Arg 195 200 205
Gly Glu Asn Gly Arg Lys Thr Arg Ile Ala Tyr Glu Arg Met Cys Asn 210
215 220 Ile Leu Lys Gly Lys Phe Gln Thr Ala Ala Gln Lys Ala Met Met
Asp 225 230 235 240 Gln Val Arg Glu Ser Arg Asp Pro Gly Asn Ala Glu
Phe Glu Asp Leu 245 250 255 Thr Phe Leu Ala Arg Ser Ala Leu Ile Leu
Arg Gly Ser Val Ala His 260 265 270 Lys Ser Cys Leu Pro Ala Cys Val
Tyr Gly Pro Ala Val Ala Ser Gly 275 280 285 Tyr Asp Phe Glu Arg Glu
Gly Tyr Ser Leu Val Gly Ile Asp Pro Phe 290 295 300 Arg Leu Leu Gln
Asn Ser Gln Val Tyr Ser Leu Ile Arg Pro Asn Glu 305 310 315 320 Asn
Pro Ala His Lys Ser 325 49252PRTInfluenza A virus 49Met Ser Leu Leu
Thr Glu Val Glu Thr Tyr Val Leu Ser Ile Ile Pro 1 5 10 15 Ser Gly
Pro Leu Lys Ala Glu Ile Ala Gln Arg Leu Glu Asp Val Phe 20 25 30
Ala Gly Lys Asn Thr Asp Leu Glu Val Leu Met Glu Trp Leu Lys Thr 35
40 45 Arg Pro Ile Leu Ser Pro Leu Thr Lys Gly Ile Leu Gly Phe Val
Phe 50 55 60 Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gln Arg Arg
Arg Phe Val 65 70 75 80 Gln Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn
Asn Met Asp Lys Ala 85 90 95 Val Lys Leu Tyr Arg Lys Leu Lys Arg
Glu Ile Thr Phe His Gly Ala 100 105 110 Lys Glu Ile Ser Leu Ser Tyr
Ser Ala Gly Ala Leu Ala Ser Cys Met 115 120 125 Gly Leu Ile Tyr Asn
Arg Met Gly Ala Val Thr Thr Glu Val Ala Phe 130 135 140 Gly Leu Val
Cys Ala Thr Cys Glu Gln Ile Ala Asp Ser Gln His Arg 145 150 155 160
Ser His Arg Gln Met Val Thr Thr Thr Asn Pro Leu Ile Arg His Glu 165
170 175 Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gln
Met 180 185 190 Ala Gly Ser Ser Glu Gln Ala Ala Glu Ala Met Glu Val
Ala Ser Gln 195 200 205 Ala Arg Gln Met Val Gln Ala Met Arg Thr Ile
Gly Thr His Pro Ser 210 215 220 Ser Ser Ala Gly Leu Lys Asn Asp Leu
Leu Glu Asn Leu Gln Ala Tyr 225 230 235 240 Gln Lys Arg Met Gly Val
Gln Met Gln Arg Phe Lys 245 250 50566PRTInfluenza A virus 50Met Lys
Ala Ile Leu Val Val Leu Leu Tyr Thr Phe Ala Thr Ala Asn 1 5 10 15
Ala Asp Thr Leu Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Asp Thr 20
25 30 Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His Ser Val
Asn 35 40 45 Leu Leu Glu Asp Lys His Asn Gly Lys Leu Cys Lys Leu
Arg Gly Val 50 55 60 Ala Pro Leu His Leu Gly Lys Cys Asn Ile Ala
Gly Trp Ile Leu Gly 65 70 75 80 Asn Pro Glu Cys Glu Ser Leu Ser Thr
Ala Ser Ser Trp Ser Tyr Ile 85 90 95 Val Glu Thr Pro Ser Ser Asp
Asn Gly Thr Cys Tyr Pro Gly Asp Phe 100 105 110 Ile Asp Tyr Glu Glu
Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe 115 120 125 Glu Arg Phe
Glu Ile Phe Pro Lys Thr Ser Ser Trp Pro Asn His Asp 130 135 140 Ser
Asn Lys Gly Val Thr Ala Ala Cys Pro His Ala Gly Ala Lys Ser 145 150
155 160 Phe Tyr Lys Asn Leu Ile Trp Leu Val Lys Lys Gly Asn Ser Tyr
Pro 165 170 175 Lys Leu Ser Lys Ser Tyr Ile Asn Asp Lys Gly Lys Glu
Val Leu Val 180 185 190 Leu Trp Gly Ile His His Pro Ser Thr Ser Ala
Asp Gln Gln Ser Leu 195 200 205 Tyr Gln Asn Ala Asp Thr Tyr Val Phe
Val Gly Ser Ser Arg Tyr Ser 210 215 220 Lys Lys Phe Lys Pro Glu Ile
Ala Ile Arg Pro Lys Val Arg Asp Gln 225 230 235 240 Glu Gly Arg Met
Asn Tyr Tyr Trp Thr Leu Val Glu Pro Gly Asp Lys 245 250 255 Ile Thr
Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg Tyr Ala Phe 260 265 270
Ala Met Glu Arg Asn Ala Gly Ser Gly Ile Ile Ile Ser Asp Thr Pro 275
280 285 Val His Asp Cys Asn Thr Thr Cys Gln Thr Pro Lys Gly Ala Ile
Asn 290 295 300 Thr Ser Leu Pro Phe Gln Asn Ile His Pro Ile Thr Ile
Gly Lys Cys 305 310 315 320 Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg
Leu Ala Thr Gly Leu Arg 325 330 335 Asn Ile Pro Ser Ile Gln Ser Arg
Gly Leu Phe Gly Ala Ile Ala Gly 340 345 350 Phe Ile Glu Gly Gly Trp
Thr Gly Met Val Asp Gly Trp Tyr Gly Tyr 355 360 365 His His Gln Asn
Glu Gln Gly Ser Gly Tyr Ala Ala Asp Leu Lys Ser 370 375 380 Thr Gln
Asn Ala Ile Asp Glu Ile Thr Asn Lys Val Asn Ser Val Ile 385 390 395
400 Glu Lys Met Asn Thr Gln Phe Thr Ala Val Gly Lys Glu Phe Asn His
405 410 415 Leu Glu Lys Arg Ile Glu Asn Leu Asn Lys Lys Val Asp Asp
Gly Phe 420 425 430 Leu Asp Ile Trp Thr Tyr Asn Ala Glu Leu Leu Val
Leu Leu Glu Asn 435 440 445 Glu Arg Thr Leu Asp Tyr His Asp Ser Asn
Val Lys Asn Leu Tyr Glu 450 455 460 Lys Val Arg Ser Gln Leu Lys Asn
Asn Ala Lys Glu Ile Gly Asn Gly 465 470 475 480 Cys Phe Glu Phe Tyr
His Lys Cys Asp Asn Thr Cys Met Glu Ser Val 485 490 495 Lys Asn Gly
Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu Ala Lys Leu 500 505 510 Asn
Arg Glu Glu Ile Asp Gly Val Lys Leu Glu Ser Thr Arg Ile Tyr 515 520
525 Gln Ile Leu Ala Ile Tyr Ser Thr Val Ala Ser Ser Leu Val Leu Val
530 535 540 Val Ser Leu Gly Ala Ile Ser Phe Trp Met Cys Ser Asn Gly
Ser Leu 545 550 555 560 Gln Cys Arg Ile Cys Ile 565
* * * * *
References